Inhibitors bases on fluoromethyl ketones. The mechanism of inhibitions of serine proteases (initially chymotrypsin) by peptidyl-fluoreomethyl ketones will be investigated. Questions of interest are: 1) Do all peptidyl-fluoromethyl ketones form adducts with the active site serine? 2) If they do, is the hydroxyl group of the resulting hemiacetal ionized? 3) What kind of interactions occur between amino-acid components of the inhibitor and the active site. These investigations will involve kinetic studies and x-ray crystallography of inhibitor enzyme complexes. This information will be used to develop strategies for the design of improved inhibitors for other serine proteases. Of primary interest is human leucocyte elastase. A new kind of active site directed inhibitor will be synthesized based on fluoromethyl ketones; targets: carboxypeptidase and angiotensin converting enzyme. Inhibitors of enzyme which utilize Schiff bases will also be synthesized, as well as inhibitors for acyl-transfer enzymes. A methionine salvage pathway. We shall continue our investigation of the pathway whereby 5'-S-methyl adenosine is converted to methionine. Intermediates and enzymes involved will be characterized. Lactyl-CoA dehydratase. An enzyme has been isolated from C1. propionicum which converts lactyl-CoA to acrylyl-CoA. The mechanism of this unusual reaction will be investigated, to determine whether a radical process is involved and what the role, if any, is of the cofactors (flavin, Fe/S centers). The composition and metabolic role of the multi-enzyme complex, of which the dehydratase is a part, will be examined.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM012633-31
Application #
2168590
Study Section
Special Emphasis Panel (NSS)
Project Start
1977-12-01
Project End
1997-11-30
Budget Start
1993-12-01
Budget End
1994-11-30
Support Year
31
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Brandeis University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
616845814
City
Waltham
State
MA
Country
United States
Zip Code
02454
Prorok, M; Albeck, A; Foxman, B M et al. (1994) Chloroketone hydrolysis by chymotrypsin and N-methylhistidyl-57-chymotrypsin: implications for the mechanism of chymotrypsin inactivation by chloroketones. Biochemistry 33:9784-90
Wray, J W; Abeles, R H (1993) A bacterial enzyme that catalyzes formation of carbon monoxide. J Biol Chem 268:21466-9
Epstein, D M; Abeles, R H (1992) Role of serine 214 and tyrosine 171, components of the S2 subsite of alpha-lytic protease, in catalysis. Biochemistry 31:11216-23
Parisi, M F; Abeles, R H (1992) Inhibition of chymotrypsin by fluorinated alpha-keto acid derivatives. Biochemistry 31:9429-35
Hu, L Y; Abeles, R H (1990) Inhibition of cathepsin B and papain by peptidyl alpha-keto esters, alpha-keto amides, alpha-diketones, and alpha-keto acids. Arch Biochem Biophys 281:271-4
Brady, K; Wei, A Z; Ringe, D et al. (1990) Structure of chymotrypsin-trifluoromethyl ketone inhibitor complexes: comparison of slowly and rapidly equilibrating inhibitors. Biochemistry 29:7600-7
Brady, K; Abeles, R H (1990) Inhibition of chymotrypsin by peptidyl trifluoromethyl ketones: determinants of slow-binding kinetics. Biochemistry 29:7608-17
Govardhan, C P; Abeles, R H (1990) Structure-activity studies of fluoroketone inhibitors of alpha-lytic protease and human leukocyte elastase. Arch Biochem Biophys 280:137-46
Abeles, R H; Alston, T A (1990) Enzyme inhibition by fluoro compounds. J Biol Chem 265:16705-8
Allen, K N; Abeles, R H (1989) Inhibition kinetics of acetylcholinesterase with fluoromethyl ketones. Biochemistry 28:8466-73

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