97 results match your criteria Applied Biological Chemistry[Journal]


Assay of β-glucosidase 2 (GBA2) activity using lithocholic acid β-3-O-glucoside substrate for cultured fibroblasts and glucosylceramide for brain tissue.

Biol Chem 2018 Dec 1. Epub 2018 Dec 1.

Department of Medical Genetics and Applied Genomics, University of Tübingen, Calwerstr. 7, D-72076 Tübingen, Germany.

Beta-glucosidase 2 (GBA2) is deficient in a form of human spastic paraplegia due to defects in the GBA2 (SPG46) gene. GBA2 was proposed as a modifier of Gaucher disease (lysosomal storage disease by deficient β-glucosidase 1; GBA1). Described assays of GBA2 activity mostly used artificial substrates incompletely modelling the situation with natural substrates in vivo. Read More

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http://dx.doi.org/10.1515/hsz-2018-0438DOI Listing
December 2018

Highly flexible, IgG-shaped, trivalent antibodies effectively target tumor cells and induce T cell-mediated killing.

Biol Chem 2019 Jan 9. Epub 2019 Jan 9.

Roche Pharma Research and Early Development, Large Molecule Research, Roche Innovation Center Munich, Nonnenwald 2, D-82377 Penzberg, Germany.

A novel bispecific antibody format was applied to generate T cell-engaging antibodies. The TriFab format is a trivalent IgG-shaped entity composed of two Fab arms that bind to antigens on the surface of tumor cells, which are linked via flexible peptides to a CD3 binding moiety that replaces the CH2 domains of conventional IgGs. The distinctive feature of these T cell recruiting bispecifics is that their CD3 variable regions are incorporated between domains, rather than N- or C-terminally fused to an Fc or antibody fragments. Read More

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http://dx.doi.org/10.1515/hsz-2018-0338DOI Listing
January 2019

Highly flexible, IgG-shaped, trivalent antibodies effectively target tumor cells and induce T cell-mediated killing.

Biol Chem 2019 Feb;400(3):343-350

Roche Pharma Research and Early Development, Large Molecule Research, Roche Innovation Center Munich, Nonnenwald 2, D-82377 Penzberg, Germany.

A novel bispecific antibody format was applied to generate T cell-engaging antibodies. The TriFab format is a trivalent IgG-shaped entity composed of two Fab arms that bind to antigens on the surface of tumor cells, which are linked via flexible peptides to a CD3 binding moiety that replaces the CH2 domains of conventional IgGs. The distinctive feature of these T cell recruiting bispecifics is that their CD3 variable regions are incorporated between domains, rather than N- or C-terminally fused to an Fc or antibody fragments. Read More

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http://dx.doi.org/10.1515/hsz-2018-0338DOI Listing
February 2019
2 Reads

Protein engineering comes of age.

Authors:
Harald Kolmar

Biol Chem 2019 Feb;400(3):255-256

Applied Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Str.-4, D-64287 Darmstadt, Germany.

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http://dx.doi.org/10.1515/hsz-2019-0108DOI Listing
February 2019

Computational design of structured loops for new protein functions.

Biol Chem 2019 Feb;400(3):275-288

Graduate Group in Biophysics, University of California San Francisco, San Francisco, CA 94158, USA.

The ability to engineer the precise geometries, fine-tuned energetics and subtle dynamics that are characteristic of functional proteins is a major unsolved challenge in the field of computational protein design. In natural proteins, functional sites exhibiting these properties often feature structured loops. However, unlike the elements of secondary structures that comprise idealized protein folds, structured loops have been difficult to design computationally. Read More

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http://dx.doi.org/10.1515/hsz-2018-0348DOI Listing
February 2019

Kinetically selective and potent inhibitors of HDAC8.

Biol Chem 2018 Dec 1. Epub 2018 Dec 1.

Department of Chemical Engineering and Biotechnology, University of Applied Sciences Darmstadt, 64295 Darmstadt, Germany.

HDAC8 is an established and validated target for T-cell lymphoma and childhood neuroblastoma. The active site binding pocket of HDAC8 is highly conserved among all zinccontaining representatives of the histone deacetylase family. This explains that most HDACs are unselectively recognized by similar inhibitors featuring a zinc binding group (ZBG), a hydrophobic linker and a head group. Read More

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http://dx.doi.org/10.1515/hsz-2018-0363DOI Listing
December 2018
7 Reads

Directed evolution of the 3C protease from coxsackievirus using a novel fluorescence-assisted intracellular method.

Biol Chem 2019 Feb;400(3):405-415

Department of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, SE-106 91 Stockholm, Sweden.

Proteases are crucial for regulating biological processes in organisms through hydrolysis of peptide bonds. Recombinant proteases have moreover become important tools in biotechnological, and biomedical research and as therapeutics. We have developed a label-free high-throughput method for quantitative assessment of proteolytic activity in Escherichia coli. Read More

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http://dx.doi.org/10.1515/hsz-2018-0362DOI Listing
February 2019
20 Reads

Microbial transglutaminase for biotechnological and biomedical engineering.

Biol Chem 2019 Feb;400(3):257-274

Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Straße 4, D-64287 Darmstadt, Germany.

Research on bacterial transglutaminase dates back to 1989, when the enzyme has been isolated from Streptomyces mobaraensis. Initially discovered during an extensive screening campaign to reduce costs in food manufacturing, it quickly appeared as a robust and versatile tool for biotechnological and pharmaceutical applications due to its excellent activity and simple handling. While pioneering attempts to make use of its extraordinary cross-linking ability resulted in heterogeneous polymers, currently it is applied to site-specifically ligate diverse biomolecules yielding precisely modified hybrid constructs comprising two or more components. Read More

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http://www.degruyter.com/view/j/bchm.ahead-of-print/hsz-2018
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http://dx.doi.org/10.1515/hsz-2018-0335DOI Listing
February 2019
15 Reads

Manipulating the stereoselectivity of a thermostable alcohol dehydrogenase by directed evolution for efficient asymmetric synthesis of arylpropanols.

Biol Chem 2019 Feb;400(3):313-321

National Engineering Laboratory for Industrial Enzymes and Tianjin Engineering Center for Biocatalytic Technology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 Xi Qi Dao, Tianjin Airport Economic Area, Tianjin 300308, China.

Chiral arylpropanols are valuable components in important pharmaceuticals and fragrances, which is the motivation for previous attempts to prepare these building blocks enantioselectively in asymmetric processes using either enzymes or transition metal catalysts. Thus far, enzymes used in kinetic resolution proved to be best, but several problems prevented ecologically and economically viable processes from being developed. In the present study, directed evolution was applied to the thermostable alcohol dehydrogenase TbSADH in the successful quest to obtain mutants that are effective in the dynamic reductive kinetic resolution (DYRKR) of racemic arylpropanals. Read More

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http://www.degruyter.com/view/j/bchm.ahead-of-print/hsz-2018
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http://dx.doi.org/10.1515/hsz-2018-0299DOI Listing
February 2019
37 Reads

A novel method for site-specific chemical SUMOylation: SUMOylation of Hsp90 modulates co-chaperone binding in vitro.

Biol Chem 2019 Mar;400(4):487-500

Department of Cell Biology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton T6G 2H7, Alberta, Canada.

SUMO is covalently attached to lysine side chains in target proteins by the action of a cascade of E1, E2, and E3 ligases. Unlike ubiquitin, SUMO does not target proteins for degradation but rather plays a regulatory role in activating target proteins or directing them to multiprotein complexes. Isolating SUMOylated proteins from native sources is challenging because of the low stoichiometry of SUMOylation that occurs for any given target protein in cells. Read More

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http://dx.doi.org/10.1515/hsz-2018-0251DOI Listing
March 2019
2 Reads

Multiple binding sites in organic cation transporters require sophisticated procedures to identify interactions of novel drugs.

Authors:
Hermann Koepsell

Biol Chem 2019 Jan;400(2):195-207

Institute of Anatomy and Cell Biology, University of Würzburg, Koellikerstr. 6, D-97070 Würzburg, Germany.

In vitro evaluation of drugs for interaction with transporters is essential during drug development. As polyspecific organic cation transporters (OCTs) are critical for pharmacokinetics of many cationic drugs, in vitro testing of human OCT1 and human OCT2 is recommended. In the currently applied tests it is determined whether uptake of one model cation in stably transfected epithelial cells is inhibited using a substrate concentration in the micromolar range. Read More

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http://dx.doi.org/10.1515/hsz-2018-0191DOI Listing
January 2019
1 Read

Biological and medical applications of plasma-activated media, water and solutions.

Biol Chem 2018 Dec;400(1):39-62

Plasma Bioscience Research Center, Applied Plasma Medicine Center, Department of Electrical and Biological Physics and Department of Plasma-Bio Display, Kwangwoon University, Seoul 01897, Republic of Korea.

Non-thermal atmospheric pressure plasma has been proposed as a new tool for various biological and medical applications. Plasma in close proximity to cell culture media or water creates reactive oxygen and nitrogen species containing solutions known as plasma-activated media (PAM) or plasma-activated water (PAW) - the latter even displays acidification. These plasma-treated solutions remain stable for several days with respect to the storage temperature. Read More

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http://dx.doi.org/10.1515/hsz-2018-0226DOI Listing
December 2018
16 Reads

The two cathepsin B-like proteases of Arabidopsis thaliana are closely related enzymes with discrete endopeptidase and carboxydipeptidase activities.

Biol Chem 2018 09;399(10):1223-1235

Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.

The genome of the model plant Arabidopsis thaliana encodes three paralogues of the papain-like cysteine proteinase cathepsin B (AtCathB1, AtCathB2 and AtCathB3), whose individual functions are still largely unknown. Here we show that a mutated splice site causes severe truncations of the AtCathB1 polypeptide, rendering it catalytically incompetent. By contrast, AtCathB2 and AtCathB3 are effective proteases which display comparable hydrolytic properties and share most of their substrate specificities. Read More

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http://dx.doi.org/10.1515/hsz-2018-0186DOI Listing
September 2018
6 Reads

Profiling system for skin kallikrein proteolysis applied in gene-deficient mouse models.

Biol Chem 2018 09;399(9):1085-1089

Laboratory of Transgenic Models of Diseases, Institute of Molecular Genetics of the ASCR, Prumyslova 595, 252 50 Vestec, Czech Republic.

Kallikrein-related proteases (KLKs) play a critical role in epidermis physiology and have been implicated in skin pathologies such as Netherton syndrome. The contribution of individual KLKs to skin proteolysis is poorly understood. Monitoring of their activities in skin proteome is hampered by overlapping substrate specificities, and there is a need for novel assays. Read More

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http://dx.doi.org/10.1515/hsz-2018-0116DOI Listing
September 2018
1 Read

Synthetic DNA filaments: from design to applications.

Biol Chem 2018 06;399(7):773-785

Centre for Medical Biotechnology (ZMB) and Centre for Nano Integration Duisburg-Essen (CENIDE), University of Duisburg-Essen, Universitätstraße 2, D-45117 Essen, Germany.

Natural filaments, such as microtubules and actin filaments, are fundamental components of the cell. Despite their relatively simple linear structure, filaments play a number of crucial roles in living organisms, from scaffolding to cellular adhesion and motility. The mechanical properties of natural filaments mostly rely on the structural features of the component units and on the way they are connected together, thus providing an ideal molecular model for emulation purposes. Read More

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http://dx.doi.org/10.1515/hsz-2018-0110DOI Listing
June 2018
3 Reads

Functional characterization of the mouse Serpina1 paralog DOM-7.

Biol Chem 2018 05;399(6):577-582

Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, D-30625 Hannover, Germany.

The generation of authentic mouse-models for human α1-antitrypsin (A1AT)-deficiency is difficult due to the high complexity of the mouse Serpina1 gene locus. Depending on the exact mouse strain, three to five paralogs are expressed, with different proteinase inhibitory properties. Nowadays with CRISPR-technology, genome editing of complex genomic loci is feasible and could be employed for the generation of A1AT-deficiency mouse models. Read More

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http://dx.doi.org/10.1515/hsz-2018-0154DOI Listing
May 2018
5 Reads

The effect of lncRNA HOTAIR on chemoresistance of ovarian cancer through regulation of HOXA7.

Biol Chem 2018 04;399(5):485-497

Department of Obstetrics and Gynecology, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, No. 32 Yihuan Road, Chengdu 610072, Sichuan, China.

This study aimed at investigating the biological functions of long non-coding RNAs (lncRNAs) hox transcript antisense intergenic RNA (HOTAIR) in resistant ovarian cancer cells, exploring the regulation effect of HOTAIR on HOXA7, and investigating their influence on the chemosensitivity of ovarian cancer cells. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied for the verification of HOTAIR expression in resistant and sensitive groups. How HOTAIR downregulation affected cell proliferation, migration and invasion, and apoptosis were determined using the MTT assay and the colony formation assay, the Transwell assay and flow cytometry analysis, respectively. Read More

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http://dx.doi.org/10.1515/hsz-2017-0274DOI Listing
April 2018
2 Reads

In vitro reconstitution and biochemical characterization of human phospholipid scramblase 3: phospholipid specificity and metal ion binding studies.

Biol Chem 2018 03;399(4):361-374

Applied and Industrial Microbiology Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600 036, India.

Human phospholipid scramblase 3 (hPLSCR3) is a single pass transmembrane protein that plays a vital role in fat metabolism, mitochondrial function, structure, maintenance and apoptosis. The mechanism of action of scramblases remains still unknown, and the role of scramblases in phospholipid translocation is heavily debated. hPLSCR3 is the only member of scramblase family localized to mitochondria and is involved in cardiolipin translocation at the mitochondrial membrane. Read More

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http://dx.doi.org/10.1515/hsz-2017-0309DOI Listing
March 2018
2 Reads

Strategies in relative and absolute quantitative mass spectrometry based proteomics.

Biol Chem 2017 05;398(5-6):687-699

Ruhr-University Bochum, Medizinisches Proteom-Center, Universitätsstraße 150, D-44801 Bochum.

Quantitative mass spectrometry approaches are used for absolute and relative quantification in global proteome studies. To date, relative and absolute quantification techniques are available that differ in quantification accuracy, proteome coverage, complexity and robustness. This review focuses on most common relative or absolute quantification strategies exemplified by three experimental studies. Read More

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http://dx.doi.org/10.1515/hsz-2017-0104DOI Listing
May 2017
2 Reads

Analysis of anticoagulants for blood-based quantitation of amyloid β oligomers in the sFIDA assay.

Biol Chem 2017 04;398(4):465-475

Early diagnostics at the preclinical stage of Alzheimer's disease is of utmost importance for drug development in clinical trials and prognostic guidance. Since soluble Aβ oligomers are considered to play a crucial role in the disease pathogenesis, several methods aim to quantify Aβ oligomers in body fluids such as cerebrospinal fluid (CSF) and blood plasma. The highly specific and sensitive method surface-based fluorescence intensity distribution analysis (sFIDA) has successfully been established for oligomer quantitation in CSF samples. Read More

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http://dx.doi.org/10.1515/hsz-2016-0153DOI Listing
April 2017
10 Reads

Genetic analysis of hereditary angioedema in a Brazilian family by targeted next generation sequencing.

Biol Chem 2016 Apr;397(4):315-22

Hereditary angioedema (HAE) is accompanied by an overproduction of bradykinin (BK) as the primary mediator of swelling. Although many proteins may be involved in regulating the wide spectrum of HAE symptoms, most studies have only focused on C1-INH and FXII. For the first time, a next generation sequencing (NGS) method was applied to develop a robust, time- and cost-effective diagnostic and research tool to analyze selected genes related to HAE. Read More

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http://dx.doi.org/10.1515/hsz-2015-0212DOI Listing
April 2016
34 Reads

Detection of thiol-based redox switch processes in parasites - facts and future.

Biol Chem 2015 May;396(5):445-63

Malaria and African trypanosomiasis are tropical diseases caused by the protozoa Plasmodium and Trypanosoma, respectively. The parasites undergo complex life cycles in the mammalian host and insect vector, during which they are exposed to oxidative and nitrosative challenges induced by the host immune system and endogenous processes. Attacking the parasite's redox metabolism is a target mechanism of several known antiparasitic drugs and a promising approach to novel drug development. Read More

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http://dx.doi.org/10.1515/hsz-2014-0279DOI Listing
May 2015
7 Reads

Subcellular distribution of FTY720 and FTY720-phosphate in immune cells - another aspect of Fingolimod action relevant for therapeutic application.

Biol Chem 2015 Jun;396(6-7):795-802

FTY720 (Fingolimod; Gilenya®) is an immune-modulatory prodrug which, after intracellular phosphorylation by sphingosine kinase 2 (SphK2) and export, mimics effects of the endogenous lipid mediator sphingosine-1-phosphate. Fingolimod has been introduced to treat relapsing-remitting multiple sclerosis. However, little has been published about the immune cell membrane penetration and subcellular distribution of FTY720 and FTY720-P. Read More

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http://dx.doi.org/10.1515/hsz-2014-0287DOI Listing
June 2015
4 Reads

Mechanisms for proteinase-activated receptor 1-triggered prostaglandin E2 generation in mouse osteoblastic MC3T3-E1 cells.

Biol Chem 2015 Feb;396(2):153-62

We analyzed signaling mechanisms for prostaglandin E2 (PGE2) production following activation of proteinase-activated receptor-1 (PAR1), a thrombin receptor, in preosteoblastic MC3T3-E1 cells. PAR1 stimulation caused PGE2 release, an effect suppressed by inhibitors of COX-1, COX-2, iPLA2, cPLA2, MAP kinases (MAPKs), Src, EGF receptor (EGFR) tyrosine kinase (EGFR-TK) and matrix metalloproteinase (MMP), but not by an intracellular Ca2+ chelator or inhibitors of PI3 kinase, protein kinase C (PKC) and NF-κB. PAR1 activation induced phosphorylation of MAPKs and upregulation of COX-2. Read More

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http://dx.doi.org/10.1515/hsz-2014-0148DOI Listing
February 2015
9 Reads

The active form of goat insulin-like peptide 3 (INSL3) is a single-chain structure comprising three domains B-C-A, constitutively expressed and secreted by testicular Leydig cells.

Biol Chem 2013 Sep;394(9):1181-94

Laboratory of Animal Reproduction and Physiology, Faculty of Agriculture, Department of Applied Biological Chemistry, Shizuoka University, Suruga-ku, Shizuoka 422-8529, Japan.

Relaxin-like factor (RLF), also called insulin-like peptide 3 (INSL3), is a member of the insulin/relaxin gene family and is produced by testicular Leydig cells. While the understanding of its effects is growing, very little is known about the structural and functional properties of native INSL3. Here, we demonstrate that native INSL3 isolated from goat testes is a single-chain structure with full biological activity, and is constitutively expressed and secreted by Leydig cells. Read More

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https://www.degruyter.com/view/j/bchm.2013.394.issue-9/hsz-2
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http://dx.doi.org/10.1515/hsz-2012-0357DOI Listing
September 2013
1 Read

Lysine-specific histone demethylase LSD1 and the dynamic control of chromatin.

Biol Chem 2013 Aug;394(8):1019-28

Institute of Biology, Developmental Genetics, Martin Luther University Halle-Wittenberg, D-06120 Halle, Germany.

The flavin adenine dinucleotide-dependent amine oxidase LSD1 is the first molecularly defined histone demethylase, which specifically demethylates H3K4me1/me2. The enzyme dynamically controls a large variety of biological processes and is associated with protein complexes controlling transcriptional repression and activation. Molecular analysis of the Drosophila LSD1 homolog revealed new insights into the epigenetic control of heterochromatin formation during early embryogenesis, the establishment of transcriptional gene silencing and the epigenetic mechanisms associated with the maintenance of stem cell identity in primordial germline cells. Read More

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http://dx.doi.org/10.1515/hsz-2013-0119DOI Listing
August 2013
13 Reads

Evaluation of the metal binding sites in a recombinant coagulation factor VIII identifies two sites with unique metal binding properties.

Biol Chem 2013 Jun;394(6):761-5

Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Måløv, Denmark.

Coagulation factor VIII is a glycosylated, non-covalent heterodimer consisting of a heavy chain (A1-A2-B domains) and a light chain (A3-C1-C2 domains). The association of the chains, and the stability and function of the dimer depend on the presence of metal ions. We applied X-ray fluorescence, X-ray crystallographic structure determination with anomalous signals at different wavelengths, and colorimetric measurements to evaluate the metal binding sites in a recombinant factor VIII molecule, turoctocog alfa. Read More

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http://dx.doi.org/10.1515/hsz-2012-0298DOI Listing
June 2013
9 Reads

Structure of the Ca(2+)-saturated C-terminal domain of scallop troponin C in complex with a troponin I fragment.

Biol Chem 2013 Jan;394(1):55-68

Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.

Troponin C (TnC) is the Ca(2+)-sensing subunit of troponin that triggers the contraction of striated muscles. In scallops, the striated muscles consume little ATP energy in sustaining strong contractile forces. The N-terminal domain of TnC works as the Ca(2+) sensor in vertebrates, whereas scallop TnC uses the C-terminal domain as the Ca(2+) sensor, suggesting that there are differences in the mechanism of the Ca(2+)-dependent regulation of muscles between invertebrates and vertebrates. Read More

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http://dx.doi.org/10.1515/hsz-2012-0152DOI Listing
January 2013
4 Reads

Biochemical and functional characterization of human phospholipid scramblase 4 (hPLSCR4).

Biol Chem 2012 Oct;393(10):1173-81

Department of Biotechnology, Applied Industrial Microbiology Laboratory, Indian Institute of Technology-Madras, Chennai 600 036, India.

Human phospholipid scramblase 4 (hPLSCR4), an isoform of the scramblase family, is a type II single-pass transmembrane protein whose function remains unknown. To understand its role, recombinant hPLSCR4 was obtained by cloning the ORF into a pET28 a(+) vector and overexpressed in Escherichia coli. Functional assay showed that Ca2+, Mg2+, and Zn2+ activate hPLSCR4 and mediate scrambling activity independent of the phospholipid head group. Read More

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http://dx.doi.org/10.1515/hsz-2012-0129DOI Listing
October 2012
1 Read

Shiga toxin glycosphingolipid receptor expression and toxin susceptibility of human pancreatic ductal adenocarcinomas of differing origin and differentiation.

Biol Chem 2012 Aug;393(8):785-99

Institute for Hygiene, University of Münster, D-48149 Münster, Germany.

Shiga toxins (Stxs) are composed of an enzymatically active A subunit (StxA) and a pentameric B subunit (StxB) that preferentially binds to the glycosphingolipid (GSL) globo\xadtriaosylceramide (Gb3Cer/CD77) and to a reduced extent to globotetraosylceramide (Gb4Cer). The identification of Gb3Cer as a tumor-associated GSL in human pancreatic cancer prompted us to investigate the expression of Gb3Cer and Gb4Cer in 15 human pancreatic ductal adenocarcinoma cell lines derived from primary tumors and liver, ascites, and lymph node metastases. Thin-layer chromatography overlay assays revealed the occurrence of Gb3Cer in all and of Gb4Cer in the majority of cell lines, which largely correlated with transcriptional expression analysis of Gb3Cer and Gb4Cer synthases. Read More

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http://dx.doi.org/10.1515/hsz-2012-0165DOI Listing
August 2012
2 Reads

Aminoquinolines as fluorescent labels for hydrophilic interaction liquid chromatography of oligosaccharides.

Biol Chem 2012 Aug;393(8):757-65

Dublin-Oxford Glycobiology Laboratory, National Institute for Bioprocessing Research and Training (NIBRT), Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland.

In this study, we investigated the potential of four different aminoquinoline (AQ) compounds as fluorescent labels for glycan analysis using hydrophilic interaction liquid chromatography (HILIC) and fluorescence detection (FLD). We confirmed the optimal excitation and emission wavelengths of 3-AQ and 6-AQ conjugated to glycan standards using three-dimensional fluorescent spectral scanning. The optimal excitation and emission wavelengths for 6-AQ were confirmed at λ(ex)=355 nm and λ(em)=440 nm. Read More

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http://dx.doi.org/10.1515/hsz-2012-0169DOI Listing
August 2012
1 Read

The analysis of N-glycans of cell membrane proteins from human hematopoietic cell lines reveals distinctions in their pattern.

Biol Chem 2012 Aug;393(8):731-47

Laboratory of Biochemistry, Department of Life Sciences and Technology, University of Applied Sciences Berlin, Seestrasse 64, 13347 Berlin, Germany.

Human cell lines are often different in their features and present variations in the glycosylation patterns of cell membrane proteins. Protein glycosylation is the most common posttranslational modification and plays a particular role in functionality and bioactivity. The key approach of this study is the comparative analysis of five hematopoietic cell lines for their N-glycosylation pattern. Read More

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https://www.degruyter.com/view/j/bchm.2012.393.issue-8/hsz-2
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http://dx.doi.org/10.1515/hsz-2012-0195DOI Listing
August 2012
13 Reads

Dye selection for live cell imaging of intact siRNA.

Biol Chem 2012 Jan;393(1-2):23-35

Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Im Neuenheimer Feld 364, D-69120 Heidelberg, Germany.

Investigations into the fate of small interfering RNA (siRNA) after transfection may unravel new ways to improve RNA interference (RNAi) efficiency. Because intracellular degradation of RNA may prevent reliable observation of fluorescence-labeled siRNA, new tools for fluorescence microscopy are warranted to cover the considerable duration of the RNAi effect. Here, the characterization and application of new fluorescence resonance energy transfer (FRET) dye pairs for sensing the integrity of duplex siRNA is reported, which allows an assessment of the degradation status of an siRNA cell population by live cell imaging. Read More

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http://dx.doi.org/10.1515/BC-2011-256DOI Listing
January 2012
5 Reads

Iron-catalyzed oxidation of Trp residues in low-density lipoprotein.

Biol Chem 2011 Oct;392(10):859-67

Section of Atherosclerosis and Lipoprotein Research, Department of Medicine, Baylor College of Medicine, Houston, TX 77030, USA.

The mechanisms of oxidation of low-density lipoproteins (LDLs) are not well defined, but epidemiological and experimental studies suggest that iron-catalyzed processes may contribute to atherogenesis. The aim of this study was to test the hypothesis that iron-catalyzed oxidations of LDLs in vitro produce diagnostic biomarkers of oxidation of the apolipoprotein that could be applied to studies in vivo. LDLs were oxidized in the presence of Fe2+, EDTA, and ascorbic acid for up to 40 h. Read More

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http://dx.doi.org/10.1515/BC.2011.173DOI Listing
October 2011
2 Reads

Improved lentiviral gene transfer into human embryonic stem cells grown in co-culture with murine feeder and stroma cells.

Biol Chem 2011 Oct 4;392(10):887-95. Epub 2011 Aug 4.

Institute for Transfusion Medicine, Hannover Medical School, D-30625 Hannover, Germany.

Genetic modification of human embryonic stem cells (hESCs) using biophysical DNA transfection methods are hampered by the very low single cell survival rate and cloning efficiency of hESCs. Lentiviral gene transfer strategies are widely used to genetically modify hESCs but limited transduction efficiencies in the presence of feeder or stroma cells present problems, particularly if vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped viral particles are applied. Here, we investigated whether the recently described semen derived enhancer of virus infection (SEVI) and alternative viral envelope proteins derived from either Gibbon ape leukaemia virus (GALV) or feline leukaemia virus (RD114) are applicable for transducing hESCs during co-culture with feeder or stroma cells. Read More

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http://dx.doi.org/10.1515/BC.2011.085DOI Listing
October 2011
7 Reads

The C2-streptavidin delivery system promotes the uptake of biotinylated molecules in macrophages and T-leukemia cells.

Biol Chem 2010 Nov;391(11):1315-25

Institute of Pharmacology and Toxicology, University of Ulm Medical Center, Albert-Einstein-Allee 11, Ulm, Germany.

Macrophages are tightly associated with inflammatory diseases as well as carcinogenesis, and therefore represent promising targets for drug delivery and gene transfer. We have recently established a novel protein delivery system based on the binary C2 toxin of Clostridium botulinum and streptavidin, allowing the uptake of exogenous biotinylated molecules into mammalian cells. Here, we applied this C2-streptavidin delivery system to macrophages and other leukocytes. Read More

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https://www.degruyter.com/view/j/bchm.2010.391.issue-11/bc.2
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http://dx.doi.org/10.1515/BC.2010.132DOI Listing
November 2010
5 Reads

Activity-based selection of HIV-1 reverse transcriptase variants with decreased polymerization fidelity.

Biol Chem 2010 Jun;391(6):665-74

Fakultät Chemie/Chemische Biologie (BCMT), Technische Universität Dortmund, Otto-Hahn-Strasse 6, D-44227 Dortmund, Germany.

HIV-1 reverse transcriptase (HIV-1 RT) copies the RNA genome of HIV-1 into DNA, thereby committing errors at an exceptionally high frequency. Viral offspring evolve rapidly and consequently are capable of evading the immune response as well as antiviral treatment. However, error-prone viral replication could drive HIV close to extinction owing to an intolerable load of deleterious mutations. Read More

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http://dx.doi.org/10.1515/BC.2010.067DOI Listing

Live cell imaging of cytoskeletal dynamics in neurons using fluorescence photoactivation.

Biol Chem 2010 Jun;391(6):639-43

Department of Neurobiology, University of Osnabrück, Barbarastrasse 11, D-49076 Osnabrück, Germany.

Alzheimer's disease and other neurodegenerative disorders share some common features at the cellular level, which are often associated with a change in cytoskeletal dynamics. Live cell imaging has been applied to study various aspects of cell physiology including cytoskeletal dynamics. Recently, fluorescence photoactivation (FPA) has been developed as a novel approach to analyze protein dynamics in living cells with high sensitivity. Read More

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http://dx.doi.org/10.1515/BC.2010.060DOI Listing

Role of N-acetyl-N-nitroso-tryptophan as nitric oxide donor in the modulation of HIF-1-dependent signaling.

Biol Chem 2010 May;391(5):533-40

Institut für Physiologie, Universität Duisburg-Essen, Germany.

N-Acetyl-N-nitroso-tryptophan (NANT) is well known for its capacity to generate nitric oxide (NO)-releasing compounds. It is unknown, however, whether NANT can be successfully applied as a precursor of NO in a complex biological environment such as a cell culture system. NO donors can be useful to induce the transcription factor hypoxia-inducible factor 1 (HIF-1) that coordinates the protection of cells and tissues from the lack of oxygen, termed hypoxia. Read More

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http://dx.doi.org/10.1515/BC.2010.054DOI Listing
May 2010
1 Read

Detection and in-cell selectivity profiling of the full-length West Nile virus NS2B/NS3 serine protease using membrane-anchored fluorescent substrates.

Biol Chem 2010 May;391(5):549-59

Department of Microbiology and Immunology, Life Sciences Center, the University of British Columbia, Vancouver, Canada.

Flaviviral NS2B/NS3 heterocomplex serine proteases are a primary target for anti-flavivirus drug discovery. To gain insights into the enzymatic properties and molecular determinants of flaviviral NS2B/NS3 protease substrate specificity in host cells, we developed and applied a novel series of membrane-anchored red-shifted fluorescent protein substrates to detect West Nile virus (WNV) NS2B/NS3 endoproteolytic activity in human cells. The substrate consists of a fluorescent reporter group (DsRed) tethered to the endoplasmic reticulum membrane by a membrane-anchoring domain. Read More

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http://dx.doi.org/10.1515/BC.2010.051DOI Listing

Identification of collagen IV derived danger/alarm signals in insect immunity by nanoLC-FTICR MS.

Biol Chem 2009 Dec;390(12):1303-11

Institute of Phytopathology and Applied Zoology, Justus Liebig University of Giessen, D-35392 Giessen, Germany.

The immune system can be stimulated by microbial molecules as well as by endogenously derived danger/alarm signals of host origin. Using the lepidopteran model insect Galleria mellonella, we recently discovered that fragments of collagen IV, resulting from hydrolysis by microbial metalloproteinases, represent danger/alarm signals in insects. Here, we characterized immune-stimulatory peptides generated by thermolysin-mediated degradation of collagen IV using nanospray ionization Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) after separation by nanoscale liquid chromatography (nanoLC). Read More

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http://www.ambiprobe.net/pdf/2009_1%20Altincicek-et-al-2009.
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http://www.degruyter.com/view/j/bchm.2009.390.issue-12/bc.20
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http://dx.doi.org/10.1515/BC.2009.128DOI Listing
December 2009

Ultrafast memory loss and relaxation processes in hydrogen-bonded systems.

Authors:
Thomas Elsaesser

Biol Chem 2009 Nov;390(11):1125-32

Max-Born-Institut für Nichtlineare Optik und Kurzzeitspektroskopie, D-12489 Berlin, Germany.

Structural memory of aqueous systems, such as neat water and biomolecules, in an aqueous environment is strongly influenced by hydrogen bond dynamics. Vibrational spectroscopy in the femtosecond (fs) time domain is applied to map structural dynamics in real-time and identify underlying molecular interactions. Neat liquid water displays an ultrafast loss of structural memory with the fastest decay of structural correlations occurring in the sub-100 fs regime. Read More

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http://dx.doi.org/10.1515/BC.2009.126DOI Listing
November 2009

Peritumoral administration of GPI-anchored TIMP-1 inhibits colon carcinoma growth in Rag-2 gamma chain-deficient mice.

Biol Chem 2009 Sep;390(9):893-7

Surgery Clinic, Technical University Munich, Munich, Germany.

Exogenous application of recombinant TIMP-1 protein modified by addition of a glycosylphosphatidylinositol (GPI) anchor allows efficient insertion of the fusion protein into cell membranes. This 'cell surface engineering' leads to changes in the proteolytic environment. TIMP-1-GPI shows enhanced as well as novel in vitro biological activities including suppression of proliferation, reduced migration, and inhibition of invasion of the colon carcinoma cell line SW480. Read More

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http://dx.doi.org/10.1515/BC.2009.098DOI Listing
September 2009
6 Reads
3 Citations
3.270 Impact Factor

On-line nano-HPLC/ESI QTOF MS monitoring of alpha2-3 and alpha2-6 sialylation in granulocyte glycosphingolipidome.

Biol Chem 2009 Jul;390(7):657-72

Institute of Medical Physics and Biophysics, University of Münster, D-48149 Münster, Germany.

A novel glycosphingolipidomic protocol using nano-high performance liquid chromatography coupled on-line to electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-QTOF-MS) focusing on the separation of isomeric ganglioside structures is described here. A highly efficient separation of alpha2-3- and alpha2-6-sialylated ganglioside species of different carbohydrate chain length was achieved on an HILIC-amido column, followed by sensitive flow-through ESI-QTOF-MS detection and unambiguous structural identification by tandem MS experiments. The protocol was applied to encompass the glycosphingolipidome of human granulocytes, where 182 distinct components could be clearly identified and assigned regarding the ganglioside type and the isomer distribution. Read More

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http://dx.doi.org/10.1515/BC.2009.066DOI Listing
July 2009
7 Reads

Metastasis-associated C4.4A, a GPI-anchored protein cleaved by ADAM10 and ADAM17.

Biol Chem 2008 Aug;389(8):1075-84

Medical Oncology Research Program, Vail d'Hebron University Hospital Research Institute, Psg. Vail d'Hebron 119-129, Universitat Autonoma de Barcelona, E-08035 Barcelona, Spain.

Metalloproteases play a complex role in tumor progression. While the activity of some ADAM, ADAMTS and matrix metalloproteases (MMPs) seems to be protumorigenic, the activity of others seems to prevent tumor progression. The identification of the array of substrates of a given metalloprotease (degradome) seems an adequate approach to predict the effect of the inhibition of a metalloprotease in tumors. Read More

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August 2008
3 Reads

Analysis of the human cytomegalovirus pp65-directed T-cell response in healthy HLA-A2-positive individuals.

Biol Chem 2008 May;389(5):551-9

InterCell AG, Campus Vienna Biocenter 6, A-1030 Vienna, Austria.

Human cytomegalovirus (HCMV) is contained by T-lymphocyte responses focused towards the major tegument protein pp65. To systematically identify T-cell epitopes, we applied the following strategy: 441 overlapping 15mer peptides spanning the entire HCMV pp65 antigen in 1-aa steps were screened in enzyme-linked immunospot (ELispot) assays for interferon gamma (IFN-gamma) secretion by peripheral blood mononuclear cells (PBMCs) from nine healthy HCMV-seropositive subjects expressing human leukocyte antigen (HLA)-A2. This analysis confirmed a number of previously known epitopes and revealed several new ones. Read More

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May 2008
4 Reads

5'-end maturation of tRNA in aquifex aeolicus.

Biol Chem 2008 Apr;389(4):395-403

Institut für Pharmazeutische Chemie, Philipps-Universität Marburg, Marbacher Weg 6, D-35037 Marburg, Germany.

5'-End maturation of tRNA primary transcripts is thought to be ubiquitously catalyzed by ribonuclease P (RNase P), a ribonucleoprotein enzyme in the vast majority of organisms and organelles. In the hyperthermophilic bacterium Aquifex aeolicus, neither a gene for the RNA nor the protein component of bacterial RNase P has been identified in its sequenced genome. Here, we demonstrate the presence of an RNase P-like activity in cell lysates of A. Read More

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http://dx.doi.org/10.1515/BC.2008.042DOI Listing
April 2008
23 Reads

Post-translational processing of selenoprotein P: implications of glycosylation for its utilisation by target cells.

Biol Chem 2007 Oct;388(10):1043-51

Institute for Biochemistry and Molecular Biology I, Heinrich Heine University Düsseldorf, Universitätstrasse 1, D-40225 Düsseldorf, Germany.

Selenoprotein P (SeP) is a highly glycosylated plasma protein containing up to 10 selenocysteine residues. It is secreted by hepatocytes and also by the human hepatoma cell line HepG2. Pharmacological inhibitors interfering with N-glycosylation, intracellular trafficking and calcium homeostasis were applied to examine post-translational processing and secretion of SeP by HepG2 cells. Read More

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https://www.degruyter.com/view/j/bchm.2007.388.issue-10/bc.2
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http://dx.doi.org/10.1515/BC.2007.136DOI Listing
October 2007
2 Reads

Alkaline Bohr effect of bird hemoglobins: the case of the flamingo.

Biol Chem 2007 Aug;388(8):787-95

Department of Applied Sciences in Biosystems, University of Cagliari, Cittadella Universitaria, I-09042 Monserrato, Italy.

The hemoglobin (Hb) substitution His-->Gln at position alpha89, very common in avian Hbs, is considered to be responsible for the weak Bohr effect of avian Hbs. Phoenicopterus ruber ruber is one of the few avian Hbs that possesses His at alpha89, but it has not been functionally characterized yet. In the present study the Hb system of the greater flamingo (P. Read More

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http://dx.doi.org/10.1515/BC.2007.102DOI Listing
August 2007
1 Read

Transgenic mouse brains for the evaluation and quality control of BSE tests.

Biol Chem 2007 Mar;388(3):349-54

European Commission, Directorate General Joint Research Centre, Institute for Reference Materials and Measurements, B-2440 Geel, Belgium.

Rapid BSE tests are widely used diagnostics in veterinary medicine and more than 11 million tests are applied worldwide. The evaluation of new rapid BSE tests and the quality assurance of approved BSE tests pose a challenge owing to the natural scarcity of BSE-infected bovine brainstems and regional variations in prion titer. Transgenic mice expressing bovine prion protein (Tg4092) offer an alternative approach to these problems. Read More

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http://dx.doi.org/10.1515/BC.2007.040DOI Listing
March 2007
8 Reads