1nff

From Proteopedia

(Difference between revisions)
Jump to: navigation, search
Line 1: Line 1:
-
[[Image:1nff.gif|left|200px]]<br /><applet load="1nff" size="350" color="white" frame="true" align="right" spinBox="true"
+
[[Image:1nff.gif|left|200px]]
-
caption="1nff, resolution 1.80&Aring;" />
+
 
-
'''Crystal structure of Rv2002 gene product from Mycobacterium tuberculosis'''<br />
+
{{Structure
 +
|PDB= 1nff |SIZE=350|CAPTION= <scene name='initialview01'>1nff</scene>, resolution 1.80&Aring;
 +
|SITE=
 +
|LIGAND= <scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</scene>
 +
|ACTIVITY= [http://en.wikipedia.org/wiki/3-alpha-(or_20-beta)-hydroxysteroid_dehydrogenase 3-alpha-(or 20-beta)-hydroxysteroid dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.53 1.1.1.53]
 +
|GENE=
 +
}}
 +
 
 +
'''Crystal structure of Rv2002 gene product from Mycobacterium tuberculosis'''
 +
 
==Overview==
==Overview==
Line 7: Line 16:
==About this Structure==
==About this Structure==
-
1NFF is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Mycobacterium_tuberculosis Mycobacterium tuberculosis] with <scene name='pdbligand=NAD:'>NAD</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/3-alpha-(or_20-beta)-hydroxysteroid_dehydrogenase 3-alpha-(or 20-beta)-hydroxysteroid dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.53 1.1.1.53] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1NFF OCA].
+
1NFF is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Mycobacterium_tuberculosis Mycobacterium tuberculosis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1NFF OCA].
==Reference==
==Reference==
-
Directed evolution approach to a structural genomics project: Rv2002 from Mycobacterium tuberculosis., Yang JK, Park MS, Waldo GS, Suh SW, Proc Natl Acad Sci U S A. 2003 Jan 21;100(2):455-60. Epub 2003 Jan 10. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=12524453 12524453]
+
Directed evolution approach to a structural genomics project: Rv2002 from Mycobacterium tuberculosis., Yang JK, Park MS, Waldo GS, Suh SW, Proc Natl Acad Sci U S A. 2003 Jan 21;100(2):455-60. Epub 2003 Jan 10. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/12524453 12524453]
[[Category: 3-alpha-(or 20-beta)-hydroxysteroid dehydrogenase]]
[[Category: 3-alpha-(or 20-beta)-hydroxysteroid dehydrogenase]]
[[Category: Mycobacterium tuberculosis]]
[[Category: Mycobacterium tuberculosis]]
Line 26: Line 35:
[[Category: psi]]
[[Category: psi]]
[[Category: sdr]]
[[Category: sdr]]
-
[[Category: structural genomics]]
+
[[Category: structural genomic]]
[[Category: tb structural genomics consortium]]
[[Category: tb structural genomics consortium]]
[[Category: tbsgc]]
[[Category: tbsgc]]
-
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:05:28 2008''
+
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 12:56:07 2008''

Revision as of 10:56, 20 March 2008

Image:1nff.gif


PDB ID 1nff

Drag the structure with the mouse to rotate
, resolution 1.80Å
Ligands:
Activity: 3-alpha-(or 20-beta)-hydroxysteroid dehydrogenase, with EC number 1.1.1.53
Coordinates: save as pdb, mmCIF, xml



Crystal structure of Rv2002 gene product from Mycobacterium tuberculosis


Overview

One of the serious bottlenecks in structural genomics projects is overexpression of the target proteins in soluble form. We have applied the directed evolution technique and prepared soluble mutants of the Mycobacterium tuberculosis Rv2002 gene product, the wild type of which had been expressed as inclusion bodies in Escherichia coli. A triple mutant I6TV47MT69K (Rv2002-M3) was chosen for structural and functional characterizations. Enzymatic assays indicate that the Rv2002-M3 protein has a high catalytic activity as a NADH-dependent 3alpha, 20beta-hydroxysteroid dehydrogenase. We have determined the crystal structures of a binary complex with NAD(+) and a ternary complex with androsterone and NADH. The structure reveals that Asp-38 determines the cofactor specificity. The catalytic site includes the triad Ser-140Tyr-153Lys-157. Additionally, it has an unusual feature, Glu-142. Enzymatic assays of the E142A mutant of Rv2002-M3 indicate that Glu-142 reverses the effect of Lys-157 in influencing the pKa of Tyr-153. This study suggests that the Rv2002 gene product is a unique member of the SDR family and is likely to be involved in steroid metabolism in M. tuberculosis. Our work demonstrates the power of the directed evolution technique as a general way of overcoming the difficulties in overexpressing the target proteins in soluble form.

About this Structure

1NFF is a Single protein structure of sequence from Mycobacterium tuberculosis. Full crystallographic information is available from OCA.

Reference

Directed evolution approach to a structural genomics project: Rv2002 from Mycobacterium tuberculosis., Yang JK, Park MS, Waldo GS, Suh SW, Proc Natl Acad Sci U S A. 2003 Jan 21;100(2):455-60. Epub 2003 Jan 10. PMID:12524453

Page seeded by OCA on Thu Mar 20 12:56:07 2008

Proteopedia Page Contributors and Editors (what is this?)

OCA

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