2puq
From Proteopedia
(New page: 200px<br /> <applet load="2puq" size="450" color="white" frame="true" align="right" spinBox="true" caption="2puq, resolution 2.050Å" /> '''Crystal structure ...) |
|||
| Line 1: | Line 1: | ||
| - | [[Image:2puq.gif|left|200px]]<br /> | + | [[Image:2puq.gif|left|200px]]<br /><applet load="2puq" size="350" color="white" frame="true" align="right" spinBox="true" |
| - | <applet load="2puq" size=" | + | |
caption="2puq, resolution 2.050Å" /> | caption="2puq, resolution 2.050Å" /> | ||
'''Crystal structure of active site inhibited coagulation factor VIIA in complex with soluble tissue factor'''<br /> | '''Crystal structure of active site inhibited coagulation factor VIIA in complex with soluble tissue factor'''<br /> | ||
| Line 6: | Line 5: | ||
==Overview== | ==Overview== | ||
The remarkably high specificity of the coagulation proteases towards, macromolecular substrates is provided by numerous interactions involving, the catalytic groove and remote exosites. For factor VIIa (FVIIa), the, principal initiator of coagulation via the extrinsic pathway, several, exosites have been identified, whereas only little is known about the, specificity dictated by the active-site architecture. Here, we have, profiled the primary P4-P1 substrate specificity of FVIIa using positional, scanning-substrate combinatorial libraries and evaluated the role of, selective active site in defining specificity. Being a trypsin-like serine, protease, FVIIa showed P1 specificity exclusively towards Arg and Lys. In, the S2 pocket Thr, Leu, Phe and Val were the most preferred amino acids., Both S3 and S4 appeared to be rather promiscuous, however, with some, preference for aromatic amino acids at both positions. Interestingly, a, significant degree of interdependence between the S3 and S4 was observed, and as a consequence, the optimal substrate for FVIIa could not be derived, directly from a sub-site directed specificity screen. To evaluate the role, of the active site residues in defining specificity, a series of mutants, of FVIIa were prepared at position 239 (c99), which is considered one of, the most important residues for determining P2 specificity of the trypsin, family members. This was confirmed for FVIIa by marked changes in primary, substrate specificity and reduced rates of antithrombin III inhibition., Interestingly, these changes do not necessarily coincide with an altered, ability to activate factor X demonstrating that inhibitor and, macromolecular substrate selectivity may be engineered separately. | The remarkably high specificity of the coagulation proteases towards, macromolecular substrates is provided by numerous interactions involving, the catalytic groove and remote exosites. For factor VIIa (FVIIa), the, principal initiator of coagulation via the extrinsic pathway, several, exosites have been identified, whereas only little is known about the, specificity dictated by the active-site architecture. Here, we have, profiled the primary P4-P1 substrate specificity of FVIIa using positional, scanning-substrate combinatorial libraries and evaluated the role of, selective active site in defining specificity. Being a trypsin-like serine, protease, FVIIa showed P1 specificity exclusively towards Arg and Lys. In, the S2 pocket Thr, Leu, Phe and Val were the most preferred amino acids., Both S3 and S4 appeared to be rather promiscuous, however, with some, preference for aromatic amino acids at both positions. Interestingly, a, significant degree of interdependence between the S3 and S4 was observed, and as a consequence, the optimal substrate for FVIIa could not be derived, directly from a sub-site directed specificity screen. To evaluate the role, of the active site residues in defining specificity, a series of mutants, of FVIIa were prepared at position 239 (c99), which is considered one of, the most important residues for determining P2 specificity of the trypsin, family members. This was confirmed for FVIIa by marked changes in primary, substrate specificity and reduced rates of antithrombin III inhibition., Interestingly, these changes do not necessarily coincide with an altered, ability to activate factor X demonstrating that inhibitor and, macromolecular substrate selectivity may be engineered separately. | ||
| - | |||
| - | ==Disease== | ||
| - | Known diseases associated with this structure: Esophageal squamous cell carcinoma OMIM:[[http://www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=606551 606551]], Factor VII deficiency OMIM:[[http://www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=227500 227500]], Myocardial infarction, decreased susceptibility to OMIM:[[http://www.ncbi.nlm.nih.gov/entrez/dispomim.cgi?id=227500 227500]] | ||
==About this Structure== | ==About this Structure== | ||
| - | 2PUQ is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with GLC, FUC, CA and CH2 as [http://en.wikipedia.org/wiki/ligands ligands]. This structure superseeds the now removed PDB entry 2PMM. Active as [http://en.wikipedia.org/wiki/Coagulation_factor_VIIa Coagulation factor VIIa], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.21 3.4.21.21] Full crystallographic information is available from [http:// | + | 2PUQ is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=GLC:'>GLC</scene>, <scene name='pdbligand=FUC:'>FUC</scene>, <scene name='pdbligand=CA:'>CA</scene> and <scene name='pdbligand=CH2:'>CH2</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. This structure superseeds the now removed PDB entry 2PMM. Active as [http://en.wikipedia.org/wiki/Coagulation_factor_VIIa Coagulation factor VIIa], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.21 3.4.21.21] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2PUQ OCA]. |
==Reference== | ==Reference== | ||
| Line 26: | Line 22: | ||
[[Category: active site inhibitor; substrate profile]] | [[Category: active site inhibitor; substrate profile]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 12:05:02 2008'' |
Revision as of 10:05, 23 January 2008
|
Crystal structure of active site inhibited coagulation factor VIIA in complex with soluble tissue factor
Overview
The remarkably high specificity of the coagulation proteases towards, macromolecular substrates is provided by numerous interactions involving, the catalytic groove and remote exosites. For factor VIIa (FVIIa), the, principal initiator of coagulation via the extrinsic pathway, several, exosites have been identified, whereas only little is known about the, specificity dictated by the active-site architecture. Here, we have, profiled the primary P4-P1 substrate specificity of FVIIa using positional, scanning-substrate combinatorial libraries and evaluated the role of, selective active site in defining specificity. Being a trypsin-like serine, protease, FVIIa showed P1 specificity exclusively towards Arg and Lys. In, the S2 pocket Thr, Leu, Phe and Val were the most preferred amino acids., Both S3 and S4 appeared to be rather promiscuous, however, with some, preference for aromatic amino acids at both positions. Interestingly, a, significant degree of interdependence between the S3 and S4 was observed, and as a consequence, the optimal substrate for FVIIa could not be derived, directly from a sub-site directed specificity screen. To evaluate the role, of the active site residues in defining specificity, a series of mutants, of FVIIa were prepared at position 239 (c99), which is considered one of, the most important residues for determining P2 specificity of the trypsin, family members. This was confirmed for FVIIa by marked changes in primary, substrate specificity and reduced rates of antithrombin III inhibition., Interestingly, these changes do not necessarily coincide with an altered, ability to activate factor X demonstrating that inhibitor and, macromolecular substrate selectivity may be engineered separately.
About this Structure
2PUQ is a Protein complex structure of sequences from Homo sapiens with , , and as ligands. This structure superseeds the now removed PDB entry 2PMM. Active as Coagulation factor VIIa, with EC number 3.4.21.21 Full crystallographic information is available from OCA.
Reference
Engineering the substrate and inhibitor specificities of human coagulation factor VIIa., Larsen KS, Ostergaard H, Bjelke JR, Olsen OH, Rasmussen HB, Christensen L, Kragelund BB, Stennicke HR, Biochem J. 2007 Apr 25;. PMID:17456045
Page seeded by OCA on Wed Jan 23 12:05:02 2008
