1sio
From Proteopedia
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'''Structure of Kumamolisin-As complexed with a covalently-bound inhibitor, AcIPF''' | '''Structure of Kumamolisin-As complexed with a covalently-bound inhibitor, AcIPF''' | ||
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[[Category: Tsuruoka, N.]] | [[Category: Tsuruoka, N.]] | ||
[[Category: Wlodawer, A.]] | [[Category: Wlodawer, A.]] | ||
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| - | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 08:44:51 2008'' | |
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Revision as of 05:44, 3 May 2008
Structure of Kumamolisin-As complexed with a covalently-bound inhibitor, AcIPF
Overview
Kumamolisin-As (previously called ScpA) is the first known example of a collagenase from the sedolisin family (MEROPS S53). This enzyme is active at low pH and in elevated temperatures. In this study that used x-ray crystallographic and biochemical methods, we investigated the structural basis of the preference of this enzyme for collagen and the importance of a glutamate residue in the unique catalytic triad (Ser(278)-Glu(78)-Asp(82)) for enzymatic activity. Crystal structures of the uninhibited enzyme and its complex with a covalently bound inhibitor, N-acetyl-isoleucyl-prolyl-phenylalaninal, showed the occurrence of a narrow S2 pocket and a groove that encompasses the active site and is rich in negative charges. Limited endoproteolysis studies of bovine type-I collagen as well as kinetic studies using peptide libraries randomized at P1 and P1', showed very strong preference for arginine at the P1 position, which correlated very well with the presence of a negatively charged residue in the S1 pocket of the enzyme. All of these features, together with those predicted through comparisons with fiddler crab collagenase, a serine peptidase, rationalize the enzyme's preference for collagen. A comparison of the Arrhenius plots of the activities of kumamolisin-As with either collagen or peptides as substrates suggests that collagen should be relaxed before proteolysis can occur. The E78H mutant, in which the catalytic triad was engineered to resemble that of subtilisin, showed only 0.01% activity of the wild-type enzyme, and its structure revealed that Ser(278), His(78), and Asp(82) do not interact with each other; thus, the canonical catalytic triad is disrupted.
About this Structure
1SIO is a Single protein structure of sequence from Alicyclobacillus sendaiensis. Full crystallographic information is available from OCA.
Reference
Crystallographic and biochemical investigations of kumamolisin-As, a serine-carboxyl peptidase with collagenase activity., Wlodawer A, Li M, Gustchina A, Tsuruoka N, Ashida M, Minakata H, Oyama H, Oda K, Nishino T, Nakayama T, J Biol Chem. 2004 May 14;279(20):21500-10. Epub 2004 Mar 10. PMID:15014068 Page seeded by OCA on Sat May 3 08:44:51 2008
