Structural (betaalpha)8 TIM barrel model of 3-hydroxy-3-methylglutaryl-coenzyme A lyase.

J Biol Chem 2003 Aug 13;278(31):29016-23. Epub 2003 May 13.

Unit of Biochemistry and Molecular Biology, International University of Catalonia, Spain.

This study describes three novel homozygous missense mutations (S75R, S201Y, and D204N) in the 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) lyase gene, which caused 3-hydroxy-3-methylglutaric aciduria in patients from Germany, England, and Argentina. Expression studies in Escherichia coli show that S75R and S201Y substitutions completely abolished the HMG-CoA lyase activity, whereas D204N reduced catalytic efficiency to 6.6% of the wild type. We also propose a three-dimensional model for human HMG-CoA lyase containing a (betaalpha)8 (TIM) barrel structure. The model is supported by the similarity with analogous TIM barrel structures of functionally related proteins, by the localization of catalytic amino acids at the active site, and by the coincidence between the shape of the substrate (HMG-CoA) and the predicted inner cavity. The three novel mutations explain the lack of HMG-CoA lyase activity on the basis of the proposed structure: in S75R and S201Y because the new amino acid residues occlude the substrate cavity, and in D204N because the mutation alters the electrochemical environment of the active site. We also report the localization of all missense mutations reported to date and show that these mutations are located in the beta-sheets around the substrate cavity.

Download full-text PDF

Source
http://dx.doi.org/10.1074/jbc.M304276200DOI Listing
August 2003

Publication Analysis

Top Keywords

hmg-coa lyase
16
s75r s201y
12
tim barrel
12
missense mutations
8
active site
8
substrate cavity
8
three novel
8
lyase activity
8
betaalpha8 tim
8
hmg-coa
5
lyase
5
catalytic amino
4
localization catalytic
4
proteins localization
4
catalytic efficiency
4
functionally proteins
4
amino acids
4
reduced catalytic
4
shape substrate
4
substrate hmg-coa
4

Similar Publications