6qd0
MloK1 model from single particle analysis of 2D crystals, class 4 (compact/open conformation)MloK1 model from single particle analysis of 2D crystals, class 4 (compact/open conformation)
Structural highlights
Function[CNGK1_RHILO] Cyclic nucleotide-regulated potassium channel activated by cAMP.[1] Publication Abstract from PubMedElectron crystallography can reveal the structure of membrane proteins within 2D crystals under close-to-native conditions. High-resolution structural information can only be reached if crystals are perfectly flat and highly ordered. In practice, such crystals are difficult to obtain. Available image unbending algorithms correct for disorder, but only perform well on images of non-tilted, flat crystals, while out-of-plane distortions are not addressed. Here, we present an approach that employs single-particle refinement procedures to locally unbend crystals in 3D. With this method, density maps of the MloK1 potassium channel with a resolution of 4 A were obtained from images of 2D crystals that do not diffract beyond 10 A. Furthermore, 3D classification allowed multiple structures to be resolved, revealing a series of MloK1 conformations within a single 2D crystal. This conformational heterogeneity explains the poor diffraction observed and is related to channel function. The approach is implemented in the FOCUS package. Retrieving high-resolution information from disordered 2D crystals by single-particle cryo-EM.,Righetto RD, Biyani N, Kowal J, Chami M, Stahlberg H Nat Commun. 2019 Apr 12;10(1):1722. doi: 10.1038/s41467-019-09661-5. PMID:30979902[2] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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