4xvp
X-ray structure of bGFP-C / EGFP complex
Structural highlights
FunctionGFP_AEQVI Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. Publication Abstract from PubMedA family of artificial proteins, named alphaRep, based on a natural family of helical repeat was previously designed. alphaRep members are efficiently expressed, folded and extremely stable proteins. A large alphaRep library was constructed creating proteins with a randomized interaction surface. In the present study, we show that the alphaRep library is an efficient source of tailor-made specific proteins with direct applications in biochemistry and cell biology. From this library, we selected by phage display alphaRep binders with nanomolar dissociation constants against the GFP. The structures of two independent alphaRep binders in complex with the GFP target were solved by X-ray crystallography revealing two totally different binding modes. The affinity of the selected alphaReps for GFP proved sufficient for practically useful applications such as pull-down experiments. alphaReps are disulfide free proteins and are efficiently and functionally expressed in eukaryotic cells: GFP-specific alphaReps are clearly sequestrated by their cognate target protein addressed to various cell compartments. These results suggest that alphaRep proteins with tailor-made specificity can be selected and used in living cells to track, modulate or interfere with intracellular processes. Specific GFP-binding artificial proteins (alphaRep): a new tool for in vitro to live cell applications.,Chevrel A, Urvoas A, de la Sierra-Gallay IL, Aumont-Nicaise M, Moutel S, Desmadril M, Perez F, Gautreau A, van Tilbeurgh H, Minard P, Valerio-Lepiniec M Biosci Rep. 2015 Jun 12;35(4). pii: e00223. doi: 10.1042/BSR20150080. PMID:26182430[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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