2AE3

Glutaryl 7-Aminocephalosporanic Acid Acylase: mutational study of activation mechanism

Summary for 2AE3

• Entry DOI: 10.2210/pdb2ae3/pdb
• Related: 2ADV 2AE4 2AE5
• Descriptor: Glutaryl 7-Aminocephalosporanic Acid Acylase, GLYCEROL, ... (4 entities in total)
• Functional Keywords: autoproteolysis, precursor activation, intermediate structure, cephalosporin acylase, hydrolase
• Biological source: Pseudomonas sp.; More
• Total number of polymer chains: 2
• Total formula weight: 77378.75

Authors

Kim, J.K.,Yang, I.S.,Shin, H.J.,Cho, K.J.,Ryu, E.K.,Kim, S.H.,Park, S.S.,Kim, K.H. (deposition date: 2005-07-21, release date: 2006-01-24, Last modification date: 2024-03-13)

Primary citation

Kim, J.K.,Yang, I.S.,Shin, H.J.,Cho, K.J.,Ryu, E.K.,Kim, S.H.,Park, S.S.,Kim, K.H.
Insight into autoproteolytic activation from the structure of cephalosporin acylase: a protein with two proteolytic chemistries.
Proc.Natl.Acad.Sci.USA, 103:1732-1737, 2006

PubMed 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.

PubMed: 16446446
DOI: 10.1073/pnas.0507862103

Experimental method

X-RAY DIFFRACTION (2.4 Å)