Structural features of cephalosporin acylase reveal the basis of autocatalytic activation

Ki Joon Cho, Jin Kwang Kim, Ji Hye Lee, Hye Jeong Shin, Sung Soo Park, Kyung Hyun Kim

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Cephalosporin acylase (CA), a member of the N-terminal nucleophile hydrolase family, is activated through two steps of intramolecular autoproteolysis, the first mediated by a serine residue, and the second by a glutamate, which releases the pro-segment and produces an active enzyme. In this study, we have determined the crystal structures of mutants which could affect primary or secondary auto-cleavage and of sequential intermediates of a slow-processing mutant at 2.0-2.5 Å resolutions. The pro-segments of the mutants undergo dynamic conformational changes during activation and adopt surprisingly different loop conformations from one another. However, the autoproteolytic site was found to form a catalytically competent conformation with a solvent water molecule, which was essentially conserved in the CA mutants.

Original languageEnglish
Pages (from-to)342-348
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume390
Issue number2
DOIs
Publication statusPublished - 2009 Dec 11

Fingerprint

Penicillin Amidase
Conformations
Chemical activation
Serine
Glutamic Acid
Crystal structure
Molecules
Water
Enzymes
Processing

Keywords

  • Autoproteolysis
  • Cephalosporin acylase
  • Intermediate structure
  • Precursor activation
  • Slow-processing mutant

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Structural features of cephalosporin acylase reveal the basis of autocatalytic activation. / Cho, Ki Joon; Kim, Jin Kwang; Lee, Ji Hye; Shin, Hye Jeong; Park, Sung Soo; Kim, Kyung Hyun.

In: Biochemical and Biophysical Research Communications, Vol. 390, No. 2, 11.12.2009, p. 342-348.

Research output: Contribution to journalArticle

Cho, Ki Joon ; Kim, Jin Kwang ; Lee, Ji Hye ; Shin, Hye Jeong ; Park, Sung Soo ; Kim, Kyung Hyun. / Structural features of cephalosporin acylase reveal the basis of autocatalytic activation. In: Biochemical and Biophysical Research Communications. 2009 ; Vol. 390, No. 2. pp. 342-348.
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