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 | |
| Publication status | Published - 2009 Dec 11 |
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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 journal › Article
}
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
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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UR - http://www.scopus.com/inward/citedby.url?scp=70350128754&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2009.09.134
DO - 10.1016/j.bbrc.2009.09.134
M3 - Article
VL - 390
SP - 342
EP - 348
JO - The BMJ
JF - The BMJ
SN - 0730-6512
IS - 2
ER -