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
References
Jan 1, 1991·Proteins·A NichollsB Honig
Mar 1, 1991·Acta Crystallographica. Section A, Foundations of Crystallography·T A JonesM Kjeldgaard
Dec 1, 1987·Journal of Bacteriology·A MatsudaK Komatsu
Nov 23, 1995·Nature·J A BranniganA G Murzin
Jun 1, 1995·European Journal of Biochemistry·Y IshiiK Shimomura
Feb 1, 1995·Protein Engineering·A C WallaceJ M Thornton
Apr 7, 1995·Journal of Molecular Biology·A G MurzinC Chothia
Jan 19, 1995·Nature·H J DugglebyP C Moody
Aug 1, 1996·Applied and Environmental Microbiology·Y SaitoK Shimomura
Aug 29, 1998·Journal of Bacteriology·Y S Lee, S S Park
May 1, 1999·Journal of Structural Biology·D E McRee
Jul 20, 1999·European Journal of Biochemistry·Y LiE Wang
Jul 31, 1999·International Journal for Parasitology·S M JayasenaE H Karunanayake
Aug 18, 2000·Journal of Structural Biology·T H KwonK H Kim
Sep 19, 2000·The Journal of Biological Chemistry·Y S LeeS S Park
Nov 18, 2000·Structure·Y KimW G Hol
Jan 1, 1997·Methods in Enzymology·E A Merritt, D J Bacon
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
Aug 27, 2009·BMC Structural Biology·Kristina LakomekTorben Lübke
Dec 19, 2013·PloS One·Matthieu TerwagneJean-Jacques Letesson
Jun 3, 2004·Chembiochem : a European Journal of Chemical Biology·Linda G OttenWim J Quax
Jun 29, 2004·Applied Microbiology and Biotechnology·Koji NagaoMitsuyoshi Ueda
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
Jan 22, 2003·Molecular Microbiology·Yi-Han LinLian-Hui Zhang
Oct 10, 2003·The Journal of Biological Chemistry·Jongchul YoonYoungsoo Kim
May 18, 2011·The Journal of Biological Chemistry·Jun YinXi Huang
Oct 10, 2007·The FEBS Journal·Linda G OttenWim J Quax