TY - JOUR
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
N1 - Funding Information:
We thank the staff at beamline 4A, Pohang Light Source for help with data collection. This work was supported by Biogreen 21 Program ( 20080401-034-008 ), Korea Research Foundation Grant ( KRF-313-C00616 ) and Korea University Grant ( 2008 ).
Copyright:
Copyright 2011 Elsevier B.V., All rights reserved.
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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U2 - 10.1016/j.bbrc.2009.09.134
DO - 10.1016/j.bbrc.2009.09.134
M3 - Article
C2 - 19800869
AN - SCOPUS:70350128754
VL - 390
SP - 342
EP - 348
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -