Insight into autoproteolytic activation from the structure of cephalosporin acylase: A protein with two proteolytic chemistries

Jin Kwang Kim, In Seok Yang, Hye Jeong Shin, Ki Joon Cho, Eui Kyung Ryu, Sun Hwa Kim, Sung Soo Park, Kyung Hyun Kim

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Cephalosporin acylase (CA), a member of the N-terminal nucleophile hydrolase family, is activated through sequential primary and secondary autoproteolytic reactions with the release of a pro segment. We have determined crystal structures of four CA mutants. Two mutants are trapped after the primary cleavage, and the other two undergo secondary cleavage slowly. These structures provide a look at pro-segment conformation during activation in N-terminal nucleophile hydrolases. The highly strained helical pro segment of precursor is transformed into a relaxed loop in the intermediates, suggesting that the relaxation of structural constraints drives the primary cleavage reaction. The secondary autoproteolytic step has been proposed to be intermolecular. However, our analysis provides evidence that CA is processed in two sequential steps of intramolecular autoproteolysis involving two distinct residues in the active site, the first a serine and the second a glutamate.

Original languageEnglish
Pages (from-to)1732-1737
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number6
DOIs
Publication statusPublished - 2006 Feb 15

Keywords

  • Autoproteolysis
  • Intermediate structure
  • Precursor activation
  • Pro segment

ASJC Scopus subject areas

  • General

Fingerprint Dive into the research topics of 'Insight into autoproteolytic activation from the structure of cephalosporin acylase: A protein with two proteolytic chemistries'. Together they form a unique fingerprint.

  • Cite this