This old version of Proteopedia is provided for student assignments while the new version is undergoing repairs. Content and edits done in this old version of Proteopedia after March 1, 2026 will eventually be lost when it is retired in about June of 2026.

Apply for new accounts at the new Proteopedia. Your logins will work in both the old and new versions.


1ijh

From Proteopedia

(Difference between revisions)
Jump to: navigation, search
Line 1: Line 1:
-
[[Image:1ijh.gif|left|200px]]
+
{{Seed}}
 +
[[Image:1ijh.png|left|200px]]
<!--
<!--
Line 9: Line 10:
{{STRUCTURE_1ijh| PDB=1ijh | SCENE= }}
{{STRUCTURE_1ijh| PDB=1ijh | SCENE= }}
-
'''CHOLESTEROL OXIDASE FROM STREPTOMYCES ASN485LEU MUTANT'''
+
===CHOLESTEROL OXIDASE FROM STREPTOMYCES ASN485LEU MUTANT===
-
==Overview==
+
<!--
-
Cholesterol oxidase catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one. An asparagine residue (Asn485) at the active site is believed to play an important role in catalysis. To test the precise role of Asn485, we mutated it to a leucine and carried out kinetic and crystallographic studies. Steady-state kinetic analysis revealed a 1300-fold decrease in the oxidation k(cat)/K(m) for the mutant enzyme whereas the k(cat)/K(m) for isomerization is only 60-fold slower. The primary kinetic isotope effect in the mutant-catalyzed reaction indicates that 3alpha-H transfer remains the rate-determining step. Measurement of the reduction potentials for the wild-type and N485L enzymes reveals a 76 mV decrease in the reduction potential of the FAD for the mutant enzyme relative to wild type. The crystal structure of the mutant, determined to 1.5 A resolution, reveals a repositioning of the side chain of Met122 near Leu485 to form a hydrophobic pocket. Furthermore, the movement of Met122 facilitates the binding of an additional water molecule, possibly mimicking the position of the equatorial hydroxyl group of the steroid substrate. The wild-type enzyme shows a novel N-H...pi interaction between the side chain of Asn485 and the pyrimidine ring of the cofactor. The loss of this interaction in the N485L mutant destabilizes the reduced flavin and accounts for the decreased reduction potential and rate of oxidation. Thus, the observed structural rearrangement of residues at the active site, as well as the kinetic data and thermodynamic data for the mutant, suggests that Asn485 is important for creating an electrostatic potential around the FAD cofactor enhancing the oxidation reaction.
+
The line below this paragraph, {{ABSTRACT_PUBMED_11705367}}, adds the Publication Abstract to the page
 +
(as it appears on PubMed at http://www.pubmed.gov), where 11705367 is the PubMed ID number.
 +
-->
 +
{{ABSTRACT_PUBMED_11705367}}
==About this Structure==
==About this Structure==
Line 28: Line 32:
[[Category: Oxidoreductase]]
[[Category: Oxidoreductase]]
[[Category: Steroid metabolism]]
[[Category: Steroid metabolism]]
-
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 20:04:11 2008''
+
 
 +
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 1 12:20:49 2008''

Revision as of 09:20, 1 July 2008

Image:1ijh.png

Template:STRUCTURE 1ijh

CHOLESTEROL OXIDASE FROM STREPTOMYCES ASN485LEU MUTANT

Template:ABSTRACT PUBMED 11705367

About this Structure

1IJH is a Single protein structure of sequence from Streptomyces sp.. Full crystallographic information is available from OCA.

Reference

The presence of a hydrogen bond between asparagine 485 and the pi system of FAD modulates the redox potential in the reaction catalyzed by cholesterol oxidase., Yin Y, Sampson NS, Vrielink A, Lario PI, Biochemistry. 2001 Nov 20;40(46):13779-87. PMID:11705367

Page seeded by OCA on Tue Jul 1 12:20:49 2008

Proteopedia Page Contributors and Editors (what is this?)

OCA

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