8g4e
Green Fluorescence Protein imaged on a cryo-EM imaging scaffoldGreen Fluorescence Protein imaged on a cryo-EM imaging scaffold
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 PubMedCryoelectron microscopy (Cryo-EM) has enabled structural determination of proteins larger than about 50 kDa, including many intractable by any other method, but it has largely failed for smaller proteins. Here, we obtain structures of small proteins by binding them to a rigid molecular scaffold based on a designed protein cage, revealing atomic details at resolutions reaching 2.9 A. We apply this system to the key cancer signaling protein KRAS (19 kDa in size), obtaining four structures of oncogenic mutational variants by cryo-EM. Importantly, a structure for the key G12C mutant bound to an inhibitor drug (AMG510) reveals significant conformational differences compared to prior data in the crystalline state. The findings highlight the promise of cryo-EM scaffolds for advancing the design of drug molecules against small therapeutic protein targets in cancer and other human diseases. Cryo-EM structure determination of small therapeutic protein targets at 3 A-resolution using a rigid imaging scaffold.,Castells-Graells R, Meador K, Arbing MA, Sawaya MR, Gee M, Cascio D, Gleave E, Debreczeni JE, Breed J, Leopold K, Patel A, Jahagirdar D, Lyons B, Subramaniam S, Phillips C, Yeates TO Proc Natl Acad Sci U S A. 2023 Sep 12;120(37):e2305494120. doi: , 10.1073/pnas.2305494120. Epub 2023 Sep 5. PMID:37669364[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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