1nff

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[[Image:1nff.gif|left|200px]]
[[Image:1nff.gif|left|200px]]
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{{Structure
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<!--
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|PDB= 1nff |SIZE=350|CAPTION= <scene name='initialview01'>1nff</scene>, resolution 1.80&Aring;
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The line below this paragraph, containing "STRUCTURE_1nff", creates the "Structure Box" on the page.
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|SITE=
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You may change the PDB parameter (which sets the PDB file loaded into the applet)
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|LIGAND= <scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</scene>
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or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
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|ACTIVITY= <span class='plainlinks'>[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] </span>
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or leave the SCENE parameter empty for the default display.
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|GENE=
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|DOMAIN=<span class='plainlinks'>[http://www.ncbi.nlm.nih.gov/Structure/cdd/cddsrv.cgi?uid=PRK12825 fabG], [http://www.ncbi.nlm.nih.gov/Structure/cdd/cddsrv.cgi?uid=PRK05653 fabG]</span>
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{{STRUCTURE_1nff| PDB=1nff | SCENE= }}
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|RELATEDENTRY=[[1nfq|1NFQ]], [[1nfr|1NFR]]
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|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1nff FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1nff OCA], [http://www.ebi.ac.uk/pdbsum/1nff PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1nff RCSB]</span>
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}}
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'''Crystal structure of Rv2002 gene product from Mycobacterium tuberculosis'''
'''Crystal structure of Rv2002 gene product from Mycobacterium tuberculosis'''
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==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://www.ncbi.nlm.nih.gov/pubmed/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]
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[[Category: 3-alpha-(or 20-beta)-hydroxysteroid dehydrogenase]]
 
[[Category: Mycobacterium tuberculosis]]
[[Category: Mycobacterium tuberculosis]]
[[Category: Single protein]]
[[Category: Single protein]]
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[[Category: Waldo, G S.]]
[[Category: Waldo, G S.]]
[[Category: Yang, J K.]]
[[Category: Yang, J K.]]
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[[Category: directed evolution]]
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[[Category: Directed evolution]]
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[[Category: gfp]]
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[[Category: Gfp]]
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[[Category: hydroxysteroid dehydrogenase]]
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[[Category: Hydroxysteroid dehydrogenase]]
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[[Category: protein structure initiative]]
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[[Category: Protein structure initiative]]
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[[Category: psi]]
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[[Category: Psi]]
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[[Category: sdr]]
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[[Category: Sdr]]
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[[Category: structural genomic]]
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[[Category: Structural genomic]]
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[[Category: tb structural genomics consortium]]
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[[Category: Tb structural genomics consortium]]
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[[Category: tbsgc]]
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[[Category: Tbsgc]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 02:28:09 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 22:29:39 2008''
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Revision as of 23:28, 2 May 2008

Image:1nff.gif

Template:STRUCTURE 1nff

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 Sat May 3 02:28:09 2008

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