1y3b: Difference between revisions

Revision as of 17:01, 21 February 2008

Crystal structure of the complex of subtilisin BPN' with chymotrypsin inhibitor 2 E60S mutant

OverviewOverview

A series of mutants of chymotrypsin inhibitor 2 (CI2), at residues involved in intramolecular interactions that shape and constrain the binding loop, were studied to determine their relative importance for inhibition of the serine protease subtilisin BPN', and for resistance of the inhibitor to proteolysis. These functional properties were investigated in tandem with the crystal structures of the mutant inhibitor-enzyme complexes. A dense hydrogen bonding network that supports the binding loop in the vicinity of the scissile bond was found to be important both for enzyme affinity and for stability to proteolysis. Structural analysis, in combination with biochemical measurements, allows differentiation of the structural components most important for resistance to proteolysis and/or binding. The most critical participating residues in the network were found to be Thr-58, Glu-60, Arg-65, and Gly-83. Glu-60 is more important for resistance to proteolysis than for binding, while Arg-65 and two other Arg residues play a greater role in binding than in resistance to proteolysis. Structural comparisons reveal a wide variety of subtle conformational changes in response to mutation, with built-in robustness in the hydrogen bond network, such that loss of one contact is compensated by other new contacts.

About this StructureAbout this Structure

1Y3B is a Protein complex structure of sequences from Bacillus amyloliquefaciens and Hordeum vulgare with , , and as ligands. Active as Subtilisin, with EC number 3.4.21.62 Full crystallographic information is available from OCA.

ReferenceReference

Role of the intramolecular hydrogen bond network in the inhibitory power of chymotrypsin inhibitor 2., Radisky ES, Lu CJ, Kwan G, Koshland DE Jr, Biochemistry. 2005 May 10;44(18):6823-30. PMID:15865427

Page seeded by OCA on Thu Feb 21 16:01:22 2008

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OCA

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-[[Image:1y3b.gif|left|200px]]<br /><applet load="1y3b" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1y3b.gif|left|200px]]<br /><applet load="1y3b" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1y3b, resolution 1.80&Aring;" />
 '''Crystal structure of the complex of subtilisin BPN' with chymotrypsin inhibitor 2 E60S mutant'''<br />
 ==Overview==
-A series of mutants of chymotrypsin inhibitor 2 (CI2), at residues, involved in intramolecular interactions that shape and constrain the, binding loop, were studied to determine their relative importance for, inhibition of the serine protease subtilisin BPN', and for resistance of, the inhibitor to proteolysis. These functional properties were, investigated in tandem with the crystal structures of the mutant, inhibitor-enzyme complexes. A dense hydrogen bonding network that supports, the binding loop in the vicinity of the scissile bond was found to be, important both for enzyme affinity and for stability to proteolysis., Structural analysis, in combination with biochemical measurements, allows, differentiation of the structural components most important for resistance, to proteolysis and/or binding. The most critical participating residues in, the network were found to be Thr-58, Glu-60, Arg-65, and Gly-83. Glu-60 is, more important for resistance to proteolysis than for binding, while, Arg-65 and two other Arg residues play a greater role in binding than in, resistance to proteolysis. Structural comparisons reveal a wide variety of, subtle conformational changes in response to mutation, with built-in, robustness in the hydrogen bond network, such that loss of one contact is, compensated by other new contacts.
+A series of mutants of chymotrypsin inhibitor 2 (CI2), at residues involved in intramolecular interactions that shape and constrain the binding loop, were studied to determine their relative importance for inhibition of the serine protease subtilisin BPN', and for resistance of the inhibitor to proteolysis. These functional properties were investigated in tandem with the crystal structures of the mutant inhibitor-enzyme complexes. A dense hydrogen bonding network that supports the binding loop in the vicinity of the scissile bond was found to be important both for enzyme affinity and for stability to proteolysis. Structural analysis, in combination with biochemical measurements, allows differentiation of the structural components most important for resistance to proteolysis and/or binding. The most critical participating residues in the network were found to be Thr-58, Glu-60, Arg-65, and Gly-83. Glu-60 is more important for resistance to proteolysis than for binding, while Arg-65 and two other Arg residues play a greater role in binding than in resistance to proteolysis. Structural comparisons reveal a wide variety of subtle conformational changes in response to mutation, with built-in robustness in the hydrogen bond network, such that loss of one contact is compensated by other new contacts.
 ==About this Structure==
-Y3B is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens] and [http://en.wikipedia.org/wiki/Hordeum_vulgare Hordeum vulgare] with CA, NA, CIT and 15P as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1Y3B OCA].
+Y3B is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens] and [http://en.wikipedia.org/wiki/Hordeum_vulgare Hordeum vulgare] with <scene name='pdbligand=CA:'>CA</scene>, <scene name='pdbligand=NA:'>NA</scene>, <scene name='pdbligand=CIT:'>CIT</scene> and <scene name='pdbligand=15P:'>15P</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1Y3B OCA].
 ==Reference==
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 [[Category: Protein complex]]
 [[Category: Subtilisin]]
-[[Category: Jr., D.E.Koshland.]]
+[[Category: Jr., D E.Koshland.]]
 [[Category: Kwan, G.]]
-[[Category: Lu, C.J.]]
+[[Category: Lu, C J.]]
-[[Category: Radisky, E.S.]]
+[[Category: Radisky, E S.]]
 [[Category: 15P]]
 [[Category: CA]]
@@ Line 25: / Line 25: @@
 [[Category: serine protease; inhibitor]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 06:32:18 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:01:22 2008''