3t0x: Difference between revisions
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< | ==Fluorogen Activating Protein M8VLA4(S55P) in complex with dimethylindole red== | ||
<StructureSection load='3t0x' size='340' side='right'caption='[[3t0x]], [[Resolution|resolution]] 1.96Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[3t0x]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3T0X OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3T0X 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]] 1.96Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=DIW:1-(3-SULFOPROPYL)-4-[(1E,3E)-3-(1,3,3-TRIMETHYL-1,3-DIHYDRO-2H-INDOL-2-YLIDENE)PROP-1-EN-1-YL]QUINOLINIUM'>DIW</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=PCA:PYROGLUTAMIC+ACID'>PCA</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=3t0x FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3t0x OCA], [https://pdbe.org/3t0x PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3t0x RCSB], [https://www.ebi.ac.uk/pdbsum/3t0x PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3t0x ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Novel fluorescent tools such as green fluorescent protein analogues and fluorogen activating proteins (FAPs) are useful in biological imaging for tracking protein dynamics in real time with a low fluorescence background. FAPs are single-chain variable fragments (scFvs) selected from a yeast surface display library that produce fluorescence upon binding a specific dye or fluorogen that is normally not fluorescent when present in solution. FAPs generally consist of human immunoglobulin variable heavy (V(H)) and variable light (V(L)) domains covalently attached via a glycine- and serine-rich linker. Previously, we determined that the yeast surface clone, V(H)-V(L) M8, could bind and activate the fluorogen dimethylindole red (DIR) but that the fluorogen activation properties were localized to the M8V(L) domain. We report here that both nuclear magnetic resonance and X-ray diffraction methods indicate the M8V(L) forms noncovalent, antiparallel homodimers that are the fluorogen activating species. The M8V(L) homodimers activate DIR by restriction of internal rotation of the bound dye. These structural results, together with directed evolution experiments with both V(H)-V(L) M8 and M8V(L), led us to rationally design tandem, covalent homodimers of M8V(L) domains joined by a flexible linker that have a high affinity for DIR and good quantum yields. | |||
A Variable Light Domain Fluorogen Activating Protein Homodimerizes To Activate Dimethylindole Red.,Senutovitch N, Stanfield RL, Bhattacharyya S, Rule GS, Wilson IA, Armitage BA, Waggoner AS, Berget PB Biochemistry. 2012 Mar 14. PMID:22390683<ref>PMID:22390683</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 3t0x" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
== | |||
== | |||
< | |||
[[Category: Homo sapiens]] | [[Category: Homo sapiens]] | ||
[[Category: Armitage | [[Category: Large Structures]] | ||
[[Category: Berget | [[Category: Armitage B]] | ||
[[Category: Bhattacharyya | [[Category: Berget P]] | ||
[[Category: Rule | [[Category: Bhattacharyya S]] | ||
[[Category: Senutovitch | [[Category: Rule G]] | ||
[[Category: Stanfield | [[Category: Senutovitch N]] | ||
[[Category: Waggoner | [[Category: Stanfield R]] | ||
[[Category: Wilson | [[Category: Waggoner AS]] | ||
[[Category: Wilson IA]] | |||
Latest revision as of 05:26, 21 November 2024
Fluorogen Activating Protein M8VLA4(S55P) in complex with dimethylindole redFluorogen Activating Protein M8VLA4(S55P) in complex with dimethylindole red
Structural highlights
Publication Abstract from PubMedNovel fluorescent tools such as green fluorescent protein analogues and fluorogen activating proteins (FAPs) are useful in biological imaging for tracking protein dynamics in real time with a low fluorescence background. FAPs are single-chain variable fragments (scFvs) selected from a yeast surface display library that produce fluorescence upon binding a specific dye or fluorogen that is normally not fluorescent when present in solution. FAPs generally consist of human immunoglobulin variable heavy (V(H)) and variable light (V(L)) domains covalently attached via a glycine- and serine-rich linker. Previously, we determined that the yeast surface clone, V(H)-V(L) M8, could bind and activate the fluorogen dimethylindole red (DIR) but that the fluorogen activation properties were localized to the M8V(L) domain. We report here that both nuclear magnetic resonance and X-ray diffraction methods indicate the M8V(L) forms noncovalent, antiparallel homodimers that are the fluorogen activating species. The M8V(L) homodimers activate DIR by restriction of internal rotation of the bound dye. These structural results, together with directed evolution experiments with both V(H)-V(L) M8 and M8V(L), led us to rationally design tandem, covalent homodimers of M8V(L) domains joined by a flexible linker that have a high affinity for DIR and good quantum yields. A Variable Light Domain Fluorogen Activating Protein Homodimerizes To Activate Dimethylindole Red.,Senutovitch N, Stanfield RL, Bhattacharyya S, Rule GS, Wilson IA, Armitage BA, Waggoner AS, Berget PB Biochemistry. 2012 Mar 14. PMID:22390683[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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