1rsu

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(New page: 200px<br /><applet load="1rsu" size="450" color="white" frame="true" align="right" spinBox="true" caption="1rsu, resolution 1.7&Aring;" /> '''COMPLEX BETWEEN STREP...)
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[[Image:1rsu.gif|left|200px]]<br /><applet load="1rsu" size="450" color="white" frame="true" align="right" spinBox="true"
 
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caption="1rsu, resolution 1.7&Aring;" />
 
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'''COMPLEX BETWEEN STREPTAVIDIN AND THE STREP-TAG II PEPTIDE'''<br />
 
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==Overview==
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==COMPLEX BETWEEN STREPTAVIDIN AND THE STREP-TAG II PEPTIDE==
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The Strep-tag is a selected nine-amino acid peptide (AWRHPQFGG) that, displays intrinsic binding affinity towards streptavidin and has been used, as an affinity tag for recombinant proteins. In order to elucidate the, molecular mechanism underlying this type of artificial protein-peptide, recognition, X-ray crystallographic analyses and binding measurements were, carried out. The crystal structure of the complex between recombinant core, streptavidin and the synthesized peptide was solved and refined at 1.7 A, resolution (space group I4(1)22; unit cell dimensions a = b = 58.3 A, c =, 176.9 A). The Strep-tag was bound at the same surface pocket where biotin, the natural ligand of streptavidin, gets complexed. The peptide backbone, exhibited 3(10)-helical conformation, with eight of the residues involved, in protein contacts. The C-terminal Gly-Gly moiety of the Strep-tag, participated in a salt bridge to Arg84 of streptavidin with its free, carboxylate group. This finding explained why the use of the Strep-tag in, fusions with recombinant proteins was restricted to their carboxyl end., Employing a synthetic peptide spot assay, the variant Strep-tag II was, screened, which did not have this limitation. The isomorphous crystal, structure of its complex with streptavidin revealed that a glutamate, side-chain provided the salt bridge in this case, with an otherwise almost, unchanged mode of binding. Affinity constants between the peptides and, streptavidin were measured by isothermal titration calorimetry. A value of, 2.7 x 10(4) M-1 was determined for the Strep-tag peptide, and slightly, tighter binding was seen when the Strep-tag was applied as part of a, bacterially produced fusion protein. This affinity is significantly, higher, compared with values previously reported for shorter, streptavidin-binding peptides, and agrees well with the remarkable, selectivity observed in recombinant protein purification applications.
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<StructureSection load='1rsu' size='340' side='right'caption='[[1rsu]], [[Resolution|resolution]] 1.70&Aring;' scene=''>
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== Structural highlights ==
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<table><tr><td colspan='2'>[[1rsu]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Streptomyces_avidinii Streptomyces avidinii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1RSU OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1RSU FirstGlance]. <br>
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</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.7&#8491;</td></tr>
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<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1rsu FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1rsu OCA], [https://pdbe.org/1rsu PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1rsu RCSB], [https://www.ebi.ac.uk/pdbsum/1rsu PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1rsu ProSAT]</span></td></tr>
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</table>
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== Function ==
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[https://www.uniprot.org/uniprot/SAV_STRAV SAV_STRAV] The biological function of streptavidin is not known. Forms a strong non-covalent specific complex with biotin (one molecule of biotin per subunit of streptavidin).
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== Evolutionary Conservation ==
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[[Image:Consurf_key_small.gif|200px|right]]
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Check<jmol>
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<jmolCheckbox>
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<scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/rs/1rsu_consurf.spt"</scriptWhenChecked>
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<scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked>
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<text>to colour the structure by Evolutionary Conservation</text>
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</jmolCheckbox>
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</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=1rsu ConSurf].
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<div style="clear:both"></div>
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==About this Structure==
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==See Also==
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1RSU is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Streptomyces_avidinii Streptomyces avidinii]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1RSU OCA].
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*[[Avidin 3D structures|Avidin 3D structures]]
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__TOC__
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==Reference==
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</StructureSection>
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Molecular interaction between the Strep-tag affinity peptide and its cognate target, streptavidin., Schmidt TG, Koepke J, Frank R, Skerra A, J Mol Biol. 1996 Feb 9;255(5):753-66. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=8636976 8636976]
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[[Category: Large Structures]]
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[[Category: Single protein]]
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[[Category: Streptomyces avidinii]]
[[Category: Streptomyces avidinii]]
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[[Category: Koepke, J.]]
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[[Category: Koepke J]]
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[[Category: Schmidt, T.]]
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[[Category: Schmidt T]]
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[[Category: Skerra, A.]]
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[[Category: Skerra A]]
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[[Category: complex (signal protein/peptide)]]
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[[Category: signal protein]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 01:50:56 2007''
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Current revision

COMPLEX BETWEEN STREPTAVIDIN AND THE STREP-TAG II PEPTIDE

PDB ID 1rsu

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