Publications by authors named "Eirini Mathioudaki"

4 Publications

  • Page 1 of 1

Expression, purification and characterization of the IcmG and IcmK proteins of the type IVB secretion system from Coxiella burnetii.

Protein Expr Purif 2021 May 11;186:105905. Epub 2021 May 11.

Division of Biochemistry, Department of Chemistry, University of Crete, GR-71003, Voutes, Greece. Electronic address:

Coxiella burnetii, the causative agent of Q fever, is an intracellular bacterial pathogen. Studies on Coxiella have shown that a type IVB secretion system (T4BSS) contributes to the establishment of the infection by transferring protein molecules. In this report, we focus on two core proteins of the Coxiella T4BSS, namely the IcmG/DotF protein (CBU_1626) and the IcmK/DotH protein (CBU_1628). Here we present a method for the recombinant expression of IcmG and IcmK in E. coli. IcmG was purified by Strep-Tactin affinity chromatography and size exclusion chromatography, while for the purification of IcmK an additional anion exchange chromatography step was introduced. The yields of the purified IcmG and IcmK proteins were 1.2 mg/L and 3 mg/L, respectively. The purified proteins showed predominant band on SDS-PAGE gel of 37 kDa for the IcmG and 40 kDa for the IcmK. Protein folding is confirmed by circular dichroism spectroscopy. The dynamic light scattering experiment indicated that IcmG and IcmK existed in a homogenous form. Further Blue native PAGE indicates the presences of a monomeric form for the IcmK and IcmG. Our work lays the basis for functional exploration and structural determination of IcmG and IcmK proteins of Coxiella's secretion system.
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http://dx.doi.org/10.1016/j.pep.2021.105905DOI Listing
May 2021

In the Search of Potential Serodiagnostic Proteins to Discriminate Between Acute and Chronic Q Fever in Humans. Some Promising Outcomes.

Front Cell Infect Microbiol 2020 18;10:557027. Epub 2020 Sep 18.

Laboratory of Biochemistry, Department of Chemistry, School of Science and Engineering, University of Crete, Heraklion, Greece.

is the agent that causes acute and chronic Q fever infections in humans. Although the isolates studied so far have shown that the two forms of the disease differ in virulence potential thus, implying a variance in their proteomic profile, the methods used do not deliver enough discriminatory capability and often, human infections may be mis-diagnosed. The current study adds further knowledge to the results that we have already published on the Coxiella outer membrane protein 1 (Com1). Herein we identified the proteins GroEL, Ybgf, OmpH, and UPF0422 as candidates for serodiagnostics of Q fever; following cloning, expression and purification they were further used as antigens in ELISA for the screening of patients' sera associated with chronic Q fever endocarditis, sera negative for phase I IgG, sera with at least one sample positive for phase I IgG and sera from patients who suffered from various rheumatic diseases. Blood donors were used as the controls. Sensitivity, specificity, positive predictive value, negative predictive value, and Cohen's kappa coefficient (κ) were calculated and we also performed binary logistic regression analysis to identify combinations of proteins with increased diagnostic yield. We found that proteins GroEL and Ybgf, together with Com1, play the most significant role in the correct diagnosis of chronic Q fever. Of these three proteins, it was shown that Com1 and GroEL present the highest sensitivity and specificity altogether. The results add to the existing knowledge that an antigen-based serodiagnostic test that will be able to correctly diagnose chronic Q fever may not be far from reality.
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http://dx.doi.org/10.3389/fcimb.2020.557027DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7531360PMC
September 2020

Com1 as a Promising Protein for the Differential Diagnosis of the Two Forms of Q Fever.

Pathogens 2019 Nov 18;8(4). Epub 2019 Nov 18.

Laboratory of Clinical Microbiology and Microbial Pathogenesis, Unit of Zoonoses, School of Medicine, University of Crete, 71110 Heraklion, Crete, Greece.

is the causative agent of acute and chronic Q fever in humans. Although the isolates studied so far showed a difference in virulence potential between those causing the two forms of the disease, implying a difference in their proteomic profile, the methods used so far to diagnose the two forms of the disease do not provide sufficient discriminatory capability, and human infections may be often misdiagnosed. The aim of the current study was to identify the outer membrane Com1 (CBU_1910) as a candidate protein for serodiagnostics of Q fever. The protein was cloned, expressed, purified, and used as an antigen in ELISA. The protein was then used for the screening of sera from patients suffering from chronic Q fever endocarditis, patients whose samples were negative for phase I immunoglobulin G (IgG), patients for whom at least one sample was positive for phase I IgG, and patients suffering from any kind of rheumatoid disease. Blood donors were used as the control group. Following statistical analysis, 92.4% (122/132) of the samples tested agreed with the negative clinical diagnosis, and 72.2% (26/36) agreed with the positive clinical diagnosis. Moreover, a significant correlation to the presence of the disease ( = 0.00) was calculated. The results support the idea that a Com1 antigen-based serodiagnostic test may be useful for differential diagnosis of chronic Q fever. Further studies are required to compare more immunogenic proteins of the bacterium against samples originating from patients suffering from different forms of the disease.
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http://dx.doi.org/10.3390/pathogens8040242DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6963606PMC
November 2019

Complete Genome Sequence of sp. Strain phDV1, an Isolate Capable of Efficient Degradation of Aromatic Hydrocarbons.

Microbiol Resour Announc 2019 Jan 10;8(2). Epub 2019 Jan 10.

Department of Chemistry, University of Crete, Heraklion, Greece.

Pseudomonas sp. strain phDV1 is a Gram-negative bacterium capable of degrading aromatic hydrocarbons. Here, we present the complete genome sequence of this strain, which consists of 4,727,682 bp, with a 62.3% G+C content and 4,574 genes. Multiple genes responsible for the degradation of aromatics are present in this strain.
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http://dx.doi.org/10.1128/MRA.01171-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6328649PMC
January 2019