1sio

From Proteopedia

(Difference between revisions)
Jump to: navigation, search
(New page: 200px<br /><applet load="1sio" size="450" color="white" frame="true" align="right" spinBox="true" caption="1sio, resolution 1.80&Aring;" /> '''Structure of Kumamol...)
Line 1: Line 1:
-
[[Image:1sio.jpg|left|200px]]<br /><applet load="1sio" size="450" color="white" frame="true" align="right" spinBox="true"
+
[[Image:1sio.jpg|left|200px]]<br /><applet load="1sio" size="350" color="white" frame="true" align="right" spinBox="true"
caption="1sio, resolution 1.80&Aring;" />
caption="1sio, resolution 1.80&Aring;" />
'''Structure of Kumamolisin-As complexed with a covalently-bound inhibitor, AcIPF'''<br />
'''Structure of Kumamolisin-As complexed with a covalently-bound inhibitor, AcIPF'''<br />
==Overview==
==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.
+
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==
==About this Structure==
-
1SIO is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Alicyclobacillus_sendaiensis Alicyclobacillus sendaiensis] with SO4, CA and ACE as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1SIO OCA].
+
1SIO is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Alicyclobacillus_sendaiensis Alicyclobacillus sendaiensis] with <scene name='pdbligand=SO4:'>SO4</scene>, <scene name='pdbligand=CA:'>CA</scene> and <scene name='pdbligand=ACE:'>ACE</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1SIO OCA].
==Reference==
==Reference==
Line 28: Line 28:
[[Category: kumamolisin-as]]
[[Category: kumamolisin-as]]
-
''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Sun Nov 25 00:34:55 2007''
+
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:01:54 2008''

Revision as of 13:01, 21 February 2008

Image:1sio.jpg

1sio, resolution 1.80Å

Drag the structure with the mouse to rotate

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 with , and as ligands. 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 Thu Feb 21 15:01:54 2008

Proteopedia Page Contributors and Editors (what is this?)

OCA

Retrieved from "http://52.214.119.220/wiki/index.php/1sio"
Personal tools