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

doi:10.1016/j.bbrc.2009.09.134

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

Honorary professors

Research output: Contribution to journal › Article › peer-review

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 language	English
Pages (from-to)	342-348
Number of pages	7
Journal	Biochemical and biophysical research communications
Volume	390
Issue number	2
DOIs	https://doi.org/10.1016/j.bbrc.2009.09.134
Publication status	Published - 2009 Dec 11

Keywords

Autoproteolysis
Cephalosporin acylase
Intermediate structure
Precursor activation
Slow-processing mutant

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "Structural features of cephalosporin acylase reveal the basis of autocatalytic activation",

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 {\AA} 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.",

keywords = "Autoproteolysis, Cephalosporin acylase, Intermediate structure, Precursor activation, Slow-processing mutant",

author = "Cho, {Ki Joon} and Kim, {Jin Kwang} and Lee, {Ji Hye} and Shin, {Hye Jeong} and Park, {Sung Soo} and Kim, {Kyung Hyun}",

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T1 - Structural features of cephalosporin acylase reveal the basis of autocatalytic activation

AU - Cho, Ki Joon

AU - Kim, Jin Kwang

AU - Lee, Ji Hye

AU - Shin, Hye Jeong

AU - Park, Sung Soo

AU - Kim, Kyung Hyun

PY - 2009/12/11

Y1 - 2009/12/11

N2 - 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.

AB - 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.

KW - Autoproteolysis

KW - Cephalosporin acylase

KW - Intermediate structure

KW - Precursor activation

KW - Slow-processing mutant

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