Substrate and inhibitor studies on proteinase 3

FEBS Letters

Dolman, Koert M.¹  Kam, Chih-Min²  Kerrigan, John E.²  Powers, James C.²  Von dem Borne, Albert E.G.Kr.¹  Goldschmeding, Roel¹ 

[1] Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Laboratory for Experimental and Clinical Immunology and Department of Hematology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands;School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA

关键词: Peptide chloromethyl ketone;    Peptide phosphonate;    Proteinase 3;    Substituted isocoumarin;    Peptide thioester;    Abu;    α-aminobutanoic acid;    Boc;    t-butyloxycarbonyl;    3-BrEtOIC;    3-(2-bromoethoxy)isocoumarin;    DCI;    3.4-dichloroisocoumarin;    DFP;    diisopropyl fluorophosphate;    3-EtOIC;    3-ethoxyisocoumarin;    HEPES;    4-(2-hydroxyethyl)-1-piperazineethane sulfonic acid;    HNE;    human neutrophil elastase;    MeO;    Methoxy;    Nva;    Norvaline;    PIPES;    piperazine-N;    N-bis(2-ethanesulfonic acid);    PMSF;    phenylmethanesulfonyl fluoride;    3-PrOIC;    3-propoxyisocoumarin;    pNA;    p-nitroanilide;    PPE;    porcine pancreatic elastase;    ONp;    p-nitrophenylester;    SBzl;    thiobenzyl ester;   

DOI  :  10.1016/0014-5793(92)80340-M

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Various amino acid and peptide thioesters were tested as substrates for human proteinase 3 and the best substrate is Boc-Ala-Ala-Nva-SBzl with a k _cal/K _m value of 1.0 × 10⁶nM^it-1s^it-1 Boc-Ala-Ala-AA-SBzl (AA = Val, Ala, or Met) are also good substrates with k _cal/K _m values of (1-4) × 10⁵ M⁻¹ s⁻¹. Substituted isocoumarins are potent inhibitors of proteinase 3 and the best inhibitors are 7-amino-4-chloro-3-(2-bromoethoxy)isocoumarin and 3.4-dichloroisocoumarin (DCI) with k _obs/[I] values of 4700 and 2600 M⁻¹ s⁻¹, respectively. Substituted isocoumarins. peptide phosphonates and chloromethyl ketones inhibited proteinase 3 less potently than human neutrophil elastase (HNE) by 1-2 orders of magnitude.

【 授权许可】

Unknown