Mevalonic acid is a key intermediate in the biosynthesis of cellular isoprenoids and sterols. Three enzymes, mevalonate kinase, phosphomevalonate kinase, and mevalonate 5-pyrophosphate decarboxylase, are required to metabolize mevalonate to the biosynthetically active isoprenoid, isopentenyl pyrophosphate. Initial work outlined for the enzymes of mevalonate metabolism focuses on mevalonate kinase, which can modulate production of isoprenoids and sterols. Perturbation of mevalonate kinase activity from its normal physiological range has been correlated with disease. In particular, mevalonic aciduria results from inherited mevalonate kinase deficiency. The proposed studies will result in the elucidation of structure/function correlations that account for mevalonate kinase catalysis and feedback regulation. This information will be useful for prediction of the consequences of mutations in the human gene that encodes this enzyme. Additionally, such data will facilitate the design of therapeutic strategies aimed at attenuating enzyme activity and potentially diminishing prenylation of cellular targets. Progress toward these long-term objectives will be generated by initial pursuit of four specific aims. (1) Recombinant forms of human and rat mevalonate kinase will be developed and characterized. The purified enzymes will be kinetically and structurally characterized and the mechanism of feedback inhibition established. (2) Using affinity labeling and sited-directed mutagenesis techniques, mevalonate kinase's active site will be identified and the function of active site amino acids assigned. (3) High resolution structures of rat and human mevalonate kinases will be determined by X-ray crystallography. (4) Human mevalonate kinase mutants will be modeled to determine the molecular basis for mevalonic aciduria.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK053766-04
Application #
6523701
Study Section
Medical Biochemistry Study Section (MEDB)
Program Officer
Mckeon, Catherine T
Project Start
1998-09-01
Project End
2004-08-31
Budget Start
2002-09-01
Budget End
2003-08-31
Support Year
4
Fiscal Year
2002
Total Cost
$243,902
Indirect Cost
Name
Medical College of Wisconsin
Department
Biochemistry
Type
Schools of Medicine
DUNS #
073134603
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
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Herdendorf, Timothy J; Miziorko, Henry M (2007) Functional evaluation of conserved basic residues in human phosphomevalonate kinase. Biochemistry 46:11780-8
Krepkiy, Dmitriy V; Miziorko, Henry M (2005) Investigation of the functional contributions of invariant serine residues in yeast mevalonate diphosphate decarboxylase. Biochemistry 44:2671-7
Voynova, Natalya E; Rios, Sandra E; Miziorko, Henry M (2004) Staphylococcus aureus mevalonate kinase: isolation and characterization of an enzyme of the isoprenoid biosynthetic pathway. J Bacteriol 186:61-7
Krepkiy, Dmitriy; Miziorko, Henry M (2004) Identification of active site residues in mevalonate diphosphate decarboxylase: implications for a family of phosphotransferases. Protein Sci 13:1875-81

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