1sib: Difference between revisions

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[[Image:1sib.gif|left|200px]]<br /><applet load="1sib" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:1sib.gif|left|200px]]<br /><applet load="1sib" size="350" color="white" frame="true" align="right" spinBox="true"

caption="1sib, resolution 2.4Å" />

'''REFINED CRYSTAL STRUCTURES OF SUBTILISIN NOVO IN COMPLEX WITH WILD-TYPE AND TWO MUTANT EGLINS. COMPARISON WITH OTHER SERINE PROTEINASE INHIBITOR COMPLEXES'''<br />

==Overview==

The crystal structures of the complexes formed between subtilisin Novo and, three inhibitors, eglin c, Arg45-eglin c and Lys53-eglin c have been, determined using molecular replacement and difference Fourier techniques, and refined at 2.4 A, 2.1 A, and 2.4 A resolution, respectively. The, mutants Arg45-eglin c and Lys53-eglin c were constructed by site-directed, mutagenesis in order to investigate the inhibitory specificity and, stability of eglin c. Arg45-eglin became a potent trypsin inhibitor, in, contrast to native eglin, which is an elastase inhibitor. This specificity, change was rationalized by comparing the structures of Arg45-eglin and, basic pancreatic trypsin inhibitor and their interactions with trypsin., The residue Arg53, which participates in a complex network of hydrogen, bonds formed between the core and the binding loop of eglin c, was, replaced with the shorter basic amino acid lysine in the mutant, Lys53-eglin. Two hydrogen bonds with Thr44, located in the binding loop, can no longer be formed but are partially restored by a water molecule, bound in the vicinity of Lys53. Eglin c in complexes with both subtilisin, Novo and subtilisin Carlsberg was crystallized in two different space, groups. Comparison of the complexes showed a rigid body rotation for the, eglin c core of 11.5 degrees with respect to the enzyme, probably caused, by different intermolecular contacts in both crystal forms.

The crystal structures of the complexes formed between subtilisin Novo and three inhibitors, eglin c, Arg45-eglin c and Lys53-eglin c have been determined using molecular replacement and difference Fourier techniques and refined at 2.4 A, 2.1 A, and 2.4 A resolution, respectively. The mutants Arg45-eglin c and Lys53-eglin c were constructed by site-directed mutagenesis in order to investigate the inhibitory specificity and stability of eglin c. Arg45-eglin became a potent trypsin inhibitor, in contrast to native eglin, which is an elastase inhibitor. This specificity change was rationalized by comparing the structures of Arg45-eglin and basic pancreatic trypsin inhibitor and their interactions with trypsin. The residue Arg53, which participates in a complex network of hydrogen bonds formed between the core and the binding loop of eglin c, was replaced with the shorter basic amino acid lysine in the mutant Lys53-eglin. Two hydrogen bonds with Thr44, located in the binding loop, can no longer be formed but are partially restored by a water molecule bound in the vicinity of Lys53. Eglin c in complexes with both subtilisin Novo and subtilisin Carlsberg was crystallized in two different space groups. Comparison of the complexes showed a rigid body rotation for the eglin c core of 11.5 degrees with respect to the enzyme, probably caused by different intermolecular contacts in both crystal forms.

==About this Structure==

1SIB is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] and [http://en.wikipedia.org/wiki/Hirudo_medicinalis Hirudo medicinalis] with CA as [http://en.wikipedia.org/wiki/ligand ligand]. 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=1SIB OCA].

1SIB is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] and [http://en.wikipedia.org/wiki/Hirudo_medicinalis Hirudo medicinalis] with <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. 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=1SIB OCA].

==Reference==

Line 15:

[[Category: Protein complex]]

[[Category: Subtilisin]]

[[Category: Gruetter, M.G.]]

[[Category: Gruetter, M G.]]

[[Category: Heinz, D.W.]]

[[Category: Heinz, D W.]]

[[Category: Priestle, J.P.]]

[[Category: Priestle, J P.]]

[[Category: CA]]

[[Category: serine protease/inhibitor complex]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Wed Nov~~ 21 02:24:~~06 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:01:50 2008''

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@@ Line 1: / Line 1: @@
-[[Image:1sib.gif|left|200px]]<br /><applet load="1sib" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1sib.gif|left|200px]]<br /><applet load="1sib" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1sib, resolution 2.4&Aring;" />
 '''REFINED CRYSTAL STRUCTURES OF SUBTILISIN NOVO IN COMPLEX WITH WILD-TYPE AND TWO MUTANT EGLINS. COMPARISON WITH OTHER SERINE PROTEINASE INHIBITOR COMPLEXES'''<br />
 ==Overview==
-The crystal structures of the complexes formed between subtilisin Novo and, three inhibitors, eglin c, Arg45-eglin c and Lys53-eglin c have been, determined using molecular replacement and difference Fourier techniques, and refined at 2.4 A, 2.1 A, and 2.4 A resolution, respectively. The, mutants Arg45-eglin c and Lys53-eglin c were constructed by site-directed, mutagenesis in order to investigate the inhibitory specificity and, stability of eglin c. Arg45-eglin became a potent trypsin inhibitor, in, contrast to native eglin, which is an elastase inhibitor. This specificity, change was rationalized by comparing the structures of Arg45-eglin and, basic pancreatic trypsin inhibitor and their interactions with trypsin., The residue Arg53, which participates in a complex network of hydrogen, bonds formed between the core and the binding loop of eglin c, was, replaced with the shorter basic amino acid lysine in the mutant, Lys53-eglin. Two hydrogen bonds with Thr44, located in the binding loop, can no longer be formed but are partially restored by a water molecule, bound in the vicinity of Lys53. Eglin c in complexes with both subtilisin, Novo and subtilisin Carlsberg was crystallized in two different space, groups. Comparison of the complexes showed a rigid body rotation for the, eglin c core of 11.5 degrees with respect to the enzyme, probably caused, by different intermolecular contacts in both crystal forms.
+The crystal structures of the complexes formed between subtilisin Novo and three inhibitors, eglin c, Arg45-eglin c and Lys53-eglin c have been determined using molecular replacement and difference Fourier techniques and refined at 2.4 A, 2.1 A, and 2.4 A resolution, respectively. The mutants Arg45-eglin c and Lys53-eglin c were constructed by site-directed mutagenesis in order to investigate the inhibitory specificity and stability of eglin c. Arg45-eglin became a potent trypsin inhibitor, in contrast to native eglin, which is an elastase inhibitor. This specificity change was rationalized by comparing the structures of Arg45-eglin and basic pancreatic trypsin inhibitor and their interactions with trypsin. The residue Arg53, which participates in a complex network of hydrogen bonds formed between the core and the binding loop of eglin c, was replaced with the shorter basic amino acid lysine in the mutant Lys53-eglin. Two hydrogen bonds with Thr44, located in the binding loop, can no longer be formed but are partially restored by a water molecule bound in the vicinity of Lys53. Eglin c in complexes with both subtilisin Novo and subtilisin Carlsberg was crystallized in two different space groups. Comparison of the complexes showed a rigid body rotation for the eglin c core of 11.5 degrees with respect to the enzyme, probably caused by different intermolecular contacts in both crystal forms.
 ==About this Structure==
-SIB is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] and [http://en.wikipedia.org/wiki/Hirudo_medicinalis Hirudo medicinalis] with CA as [http://en.wikipedia.org/wiki/ligand ligand]. 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=1SIB OCA].
+SIB is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] and [http://en.wikipedia.org/wiki/Hirudo_medicinalis Hirudo medicinalis] with <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. 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=1SIB OCA].
 ==Reference==
@@ Line 15: / Line 15: @@
 [[Category: Protein complex]]
 [[Category: Subtilisin]]
-[[Category: Gruetter, M.G.]]
+[[Category: Gruetter, M G.]]
-[[Category: Heinz, D.W.]]
+[[Category: Heinz, D W.]]
-[[Category: Priestle, J.P.]]
+[[Category: Priestle, J P.]]
 [[Category: CA]]
 [[Category: serine protease/inhibitor complex]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 02:24:06 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:01:50 2008''