5cp4: Difference between revisions
No edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
[[Image:5cp4.gif|left|200px]]<br /><applet load="5cp4" size=" | [[Image:5cp4.gif|left|200px]]<br /><applet load="5cp4" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="5cp4, resolution 1.75Å" /> | caption="5cp4, resolution 1.75Å" /> | ||
'''CRYOGENIC STRUCTURE OF P450CAM'''<br /> | '''CRYOGENIC STRUCTURE OF P450CAM'''<br /> | ||
| Line 7: | Line 7: | ||
==About this Structure== | ==About this Structure== | ||
5CP4 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida] with K, HEM, CAM and GOL as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Camphor_5-monooxygenase Camphor 5-monooxygenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.14.15.1 1.14.15.1] Known structural/functional Site: <scene name='pdbsite=K:K Binding Site'>K</scene>. Full crystallographic information is available from [http:// | 5CP4 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida] with <scene name='pdbligand=K:'>K</scene>, <scene name='pdbligand=HEM:'>HEM</scene>, <scene name='pdbligand=CAM:'>CAM</scene> and <scene name='pdbligand=GOL:'>GOL</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Camphor_5-monooxygenase Camphor 5-monooxygenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.14.15.1 1.14.15.1] Known structural/functional Site: <scene name='pdbsite=K:K+Binding+Site'>K</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5CP4 OCA]. | ||
==Reference== | ==Reference== | ||
| Line 26: | Line 26: | ||
[[Category: p450]] | [[Category: p450]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Feb 3 10:53:20 2008'' | ||
Revision as of 11:53, 3 February 2008
|
CRYOGENIC STRUCTURE OF P450CAM
OverviewOverview
Proton transfer in cytochromes P450 is a critical step in the activation, of molecular oxygen. Extensive study of the P450cam active site has, identified several residues that play a central role in dioxygen bond, scission. A highly conserved carboxylate, aspartate-251 in P450cam in the, distal helix I, participates in a series of hydrogen-bond/ion pairs near, the molecular surface and has been implicated in the catalytic mechanism., Mutation of Asp251 is known to lower activity by 2 orders of magnitude and, change the rate-limiting step in the catalytic cycle, suggesting a role, for an acid functionality in generation of iron-oxygen reactive, intermediates. The turnover rates of the Asp251Asn mutant in various, protium-deuterium mixtures have been determined and show a significantly, larger kinetic solvent isotope effect, with an overall magnitude of 10, compared to 1.8 for the wild-type P450cam. In addition, a much larger, number of protons are involved in the rate-limiting step for the Asp251Asn, mutant than in the wild-type enzyme. These results indicate that Asp251 is, an essential part of the normal proton delivery machinery required for O-O, bond scission. The crystal structure of the Aps251Asn mutant obtained from, data collected at cryogenic temperatures has been refined to 1.9 A. Key, hydrogen bonds required to hold Asp251 in position have been broken which, allows the mutant Asn251 side chain to swing out and away from the O2, binding site leading to a more open active site. This change could allow, easier access by water and thus contribute to the observed kinetic solvent, isotope effects.
About this StructureAbout this Structure
5CP4 is a Single protein structure of sequence from Pseudomonas putida with , , and as ligands. Active as Camphor 5-monooxygenase, with EC number 1.14.15.1 Known structural/functional Site: . Full crystallographic information is available from OCA.
ReferenceReference
Understanding the role of the essential Asp251 in cytochrome p450cam using site-directed mutagenesis, crystallography, and kinetic solvent isotope effect., Vidakovic M, Sligar SG, Li H, Poulos TL, Biochemistry. 1998 Jun 30;37(26):9211-9. PMID:9649301
Page seeded by OCA on Sun Feb 3 10:53:20 2008