Kinetic dissection of α1-antitrypsin inhibition mechanism

Jong Shik Shin, Myeong Hee Yu

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Serpins (serine protease inhibitors) inhibit target proteases by forming a stable covalent complex in which the cleaved reactive site loop of the serpin is inserted into β-sheet A of the serpin with concomitant translocation of the protease to the opposite of the initial binding site. Despite recent determination of the crystal structures of a Michaelis protease-serpin complex as well as a stable covalent complex, details on the kinetic mechanism remain unsolved mainly due to difficulties in measuring kinetic parameters of acylation, protease translocation, and deacylation steps. To address the problem, we applied a mathematical model developed on the basis of a suicide inhibition mechanism to the stopped-flow kinetics of fluorescence resonance energy transfer during complex formation between α1-antitrypsin, a prototype serpin, and proteases. Compared with the hydrolysis of a peptide substrate, acylation of the protease by α1-antitrypsin is facilitated, whereas deacylation of the acyl intermediate is strongly suppressed during the protease translocation. The results from nucleophile susceptibility of the acyl intermediate suggest strongly that the active site of the protease is already perturbed during translocation.

Original languageEnglish
Pages (from-to)11629-11635
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number14
DOIs
Publication statusPublished - 2002 Apr 5

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Dissection
Serine Proteinase Inhibitors
Peptide Hydrolases
Kinetics
Acylation
Catalytic Domain
Nucleophiles
Fluorescence Resonance Energy Transfer
Inhibition (Psychology)
Kinetic parameters
Hydrolysis
Suicide
Crystal structure
Binding Sites
Theoretical Models
Mathematical models
Peptides
Substrates

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Shin, Jong Shik ; Yu, Myeong Hee. / Kinetic dissection of α1-antitrypsin inhibition mechanism. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 14. pp. 11629-11635.
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Kinetic dissection of α1-antitrypsin inhibition mechanism. / Shin, Jong Shik; Yu, Myeong Hee.

In: Journal of Biological Chemistry, Vol. 277, No. 14, 05.04.2002, p. 11629-11635.

Research output: Contribution to journalArticle

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