1oss: Difference between revisions

New page: left|200px<br /><applet load="1oss" size="450" color="white" frame="true" align="right" spinBox="true" caption="1oss, resolution 1.93Å" /> '''T190P STREPTOMYCES G...
 
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[[Image:1oss.gif|left|200px]]<br /><applet load="1oss" size="450" color="white" frame="true" align="right" spinBox="true"
caption="1oss, resolution 1.93&Aring;" />
'''T190P STREPTOMYCES GRISEUS TRYPSIN IN COMPLEX WITH BENZAMIDINE'''<br />


==Overview==
==T190P STREPTOMYCES GRISEUS TRYPSIN IN COMPLEX WITH BENZAMIDINE==
Streptomyces griseus trypsin (SGT) was chosen as a model scaffold for the, development of serine proteases with enhanced substrate specificity., Recombinant SGT has been produced in a Bacillus subtilis expression system, in a soluble active form and purified to homogeneity. The recombinant and, native proteases have nearly identical enzymatic properties and, structures. Four SGT mutants with alterations in the S1 substrate binding, pocket (T190A, T190P, T190S, and T190V) were also expressed. The T190P, mutant demonstrated the largest shift to a preference for Arg versus Lys, in the P1 site. This was shown by a minor reduction in catalytic activity, toward an Arg-containing substrate (k(cat) reduction of 25%). The crystal, structures of the recombinant SGT and the T190P mutant in a complex with, the inhibitor benzamidine were obtained at high resolution (approximately, 1.9 A). The increase in P1 specificity, achieved with minimal effect on, the catalytic efficiency, demonstrates that the T190P mutant is an ideal, candidate for the design of additional substrate specificity engineered, into the S2 to S4 binding pockets.
<StructureSection load='1oss' size='340' side='right'caption='[[1oss]], [[Resolution|resolution]] 1.93&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[1oss]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Streptomyces_griseus Streptomyces griseus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1OSS OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1OSS FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.93&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=BEN:BENZAMIDINE'>BEN</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr>
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1oss FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1oss OCA], [https://pdbe.org/1oss PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1oss RCSB], [https://www.ebi.ac.uk/pdbsum/1oss PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1oss ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/TRYP_STRGR TRYP_STRGR]
== Evolutionary Conservation ==
[[Image:Consurf_key_small.gif|200px|right]]
Check<jmol>
  <jmolCheckbox>
    <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/os/1oss_consurf.spt"</scriptWhenChecked>
    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.spt</scriptWhenUnchecked>
    <text>to colour the structure by Evolutionary Conservation</text>
  </jmolCheckbox>
</jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1oss ConSurf].
<div style="clear:both"></div>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
Streptomyces griseus trypsin (SGT) was chosen as a model scaffold for the development of serine proteases with enhanced substrate specificity. Recombinant SGT has been produced in a Bacillus subtilis expression system in a soluble active form and purified to homogeneity. The recombinant and native proteases have nearly identical enzymatic properties and structures. Four SGT mutants with alterations in the S1 substrate binding pocket (T190A, T190P, T190S, and T190V) were also expressed. The T190P mutant demonstrated the largest shift to a preference for Arg versus Lys in the P1 site. This was shown by a minor reduction in catalytic activity toward an Arg-containing substrate (k(cat) reduction of 25%). The crystal structures of the recombinant SGT and the T190P mutant in a complex with the inhibitor benzamidine were obtained at high resolution (approximately 1.9 A). The increase in P1 specificity, achieved with minimal effect on the catalytic efficiency, demonstrates that the T190P mutant is an ideal candidate for the design of additional substrate specificity engineered into the S2 to S4 binding pockets.


==About this Structure==
Engineering the primary substrate specificity of Streptomyces griseus trypsin.,Page MJ, Wong SL, Hewitt J, Strynadka NC, MacGillivray RT Biochemistry. 2003 Aug 5;42(30):9060-6. PMID:12885239<ref>PMID:12885239</ref>
1OSS is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_chryseus Streptomyces chryseus] with SO4, CA and BEN as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Trypsin Trypsin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.4 3.4.21.4] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1OSS OCA].


==Reference==
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
Engineering the primary substrate specificity of Streptomyces griseus trypsin., Page MJ, Wong SL, Hewitt J, Strynadka NC, MacGillivray RT, Biochemistry. 2003 Aug 5;42(30):9060-6. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=12885239 12885239]
</div>
[[Category: Single protein]]
<div class="pdbe-citations 1oss" style="background-color:#fffaf0;"></div>
[[Category: Streptomyces chryseus]]
[[Category: Trypsin]]
[[Category: Hewitt, J.]]
[[Category: MacGillivray, R.T.]]
[[Category: Page, M.J.]]
[[Category: Strynadka, N.C.]]
[[Category: Wong, S.L.]]
[[Category: BEN]]
[[Category: CA]]
[[Category: SO4]]
[[Category: mutant]]
[[Category: serine protease]]
[[Category: trypsin]]


''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 23:09:46 2007''
==See Also==
*[[Trypsin 3D structures|Trypsin 3D structures]]
== References ==
<references/>
__TOC__
</StructureSection>
[[Category: Large Structures]]
[[Category: Streptomyces griseus]]
[[Category: Hewitt J]]
[[Category: MacGillivray RT]]
[[Category: Page MJ]]
[[Category: Strynadka NC]]
[[Category: Wong SL]]

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