Enzymic Paths and Mechanisms in Amino Acid Metabolism

Dekker, Eugene E.

Abstract

Research will be continued on the structure and mechanism of action of 2-keto-4-hydroxyglutarate aldolase, a key enzyme involved in the metabolism of L-hydroxyproline by mammals. Our efforts include an exploration of the functional aminoacyl groups that participate in the catalytic process, isolation of active-site peptides, and general establishment of structure-function relationships. Since we have the enzyme in pure form from extracts of bovine liver, bovine kidney, and E. coli, comparative enzymology is done. Studies will be initiated, in vivo and in vitro, on the metabolism and enzymology of gamma-methyleneglutamic acid. Further efforts will be made to purify and characterize D-1-amino-2-propanol:NADO oxidoreductase; the mechanism of action of L-threonine dehydrogenase will be examined. Both of these enzymes are found in animals and bacteria and are involved in the catabolism of L-threonine.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
• Type: Research Project (R01)
• Project #: 5R01DK003718-27
• Application #: 3224408
• Study Section: Physiological Chemistry Study Section (PC)
• Project Start: 1977-12-01
• Project End: 1987-11-30
• Budget Start: 1985-12-01
• Budget End: 1986-11-30
• Support Year: 27
• Fiscal Year: 1986

Institution

• Name: University of Michigan Ann Arbor
• Type: Schools of Medicine
• DUNS #: 791277940
• City: Ann Arbor
• State: MI
• Country: United States
• Zip Code: 48109

Publications

Chen, Y W; Dekker, E E; Somerville, R L (1995) Functional analysis of E. coli threonine dehydrogenase by means of mutant isolation and characterization. Biochim Biophys Acta 1253:208-14

Marcus, J P; Dekker, E E (1995) Identification of a second active site residue in Escherichia coli L-threonine dehydrogenase: methylation of histidine-90 with methyl p-nitrobenzenesulfonate. Arch Biochem Biophys 316:413-20

Marcus, J P; Dekker, E E (1993) pH-dependent decarboxylation of 2-amino-3-ketobutyrate, the unstable intermediate in the threonine dehydrogenase-initiated pathway for threonine utilization. Biochem Biophys Res Commun 190:1066-72

Marcus, J P; Dekker, E E (1993) Threonine formation via the coupled activity of 2-amino-3-ketobutyrate coenzyme A lyase and threonine dehydrogenase. J Bacteriol 175:6505-11

Marcus, J P; Dekker, E E (1993) Identity and some properties of the L-threonine aldolase activity manifested by pure 2-amino-3-ketobutyrate ligase of Escherichia coli. Biochim Biophys Acta 1164:299-304

Mukherjee, J J; Dekker, E E (1992) Inactivation of Escherichia coli 2-amino-3-ketobutyrate CoA ligase by phenylglyoxal and identification of an active-site arginine peptide. Arch Biochem Biophys 299:147-53

Patil, R V; Dekker, E E (1992) Cloning, nucleotide sequence, overexpression, and inactivation of the Escherichia coli 2-keto-4-hydroxyglutarate aldolase gene. J Bacteriol 174:102-7

Dekker, E E; Kitson, R P (1992) 2-Keto-4-hydroxyglutarate aldolase: purification and characterization of the homogeneous enzyme from bovine kidney. J Biol Chem 267:10507-14

Epperly, B R; Dekker, E E (1991) L-threonine dehydrogenase from Escherichia coli. Identification of an active site cysteine residue and metal ion studies. J Biol Chem 266:6086-92

Mukherjee, J J; Dekker, E E (1990) 2-Amino-3-ketobutyrate CoA ligase of Escherichia coli: stoichiometry of pyridoxal phosphate binding and location of the pyridoxyllysine peptide in the primary structure of the enzyme. Biochim Biophys Acta 1037:24-9