Active site residues of cephalosporin acylase are critical not only for enzymatic catalysis but also for post-translational modification

The Journal of Biological Chemistry
S Kim, Y Kim

Abstract

Cephalosporin acylase (CA) is a recently identified N-terminal hydrolase. It is also a commercially important enzyme in producing 7-aminocephalosporanic acid (7-ACA), a backbone chemical in synthesizing semi-synthetic cephalosporin antibiotics. CA is translated as an inactive single chain precursor, being post-translationally modified into an active enzyme. The post-translational modification takes place in two steps. The first intramolecular autocatalytic proteolysis takes place at one end of the spacer peptide by a nucleophilic Ser or Thr, which in turn becomes a new N-terminal Ser or Thr. The second intermolecular modification cleaves off the other end of the spacer peptide by another CA. Two binary structures in complex with glutaryl-7-ACA (the most favored substrate of CAs) and glutarate (side chain of glutaryl-7-ACA) were determined, and they revealed the detailed interactions of glutaryl-7-ACA with the active site residues (Y. Kim and W. G. J. Hol (2001) Chem. Biol., in press). In this report: 1) we have mutated key active site residues into nonfunctional amino acids, and their roles in catalysis were further analyzed; 2) we performed mutagenesis studies indicating that secondary intermolecular modification is carried ou...Continue Reading

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Citations

Feb 1, 2006·Proceedings of the National Academy of Sciences of the United States of America·Jin Kwang KimKyung Hyun Kim
Sep 10, 2005·Applied and Environmental Microbiology·Wei ZhangWeihong Jiang
Feb 24, 2006·Infection and Immunity·Charles F SioWim J Quax
Dec 19, 2013·PloS One·Matthieu TerwagneJean-Jacques Letesson
Jun 3, 2004·Chembiochem : a European Journal of Chemical Biology·Linda G OttenWim J Quax
Sep 19, 2002·The Journal of Biological Chemistry·Hideyuki Suzuki, Hidehiko Kumagai
Jul 1, 2006·Critical Reviews in Biotechnology·Vijay Chintaman Sonawane
Sep 17, 2002·European Journal of Biochemistry·Charles F SioWim J Quax
Oct 10, 2003·The Journal of Biological Chemistry·Jongchul YoonYoungsoo Kim
Oct 10, 2007·The FEBS Journal·Linda G OttenWim J Quax

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