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• W2732605882 endingPage "2508" @default.
• W2732605882 startingPage "2490" @default.
• W2732605882 abstract "The actin cytoskeleton is a complex network controlled by a vast array of intricately regulated actin-binding proteins. Human plastins (PLS1, PLS2, and PLS3) are evolutionary conserved proteins that non-covalently crosslink actin filaments into tight bundles. Through stabilization of such bundles, plastins contribute, in an isoform-specific manner, to the formation of kidney and intestinal microvilli, inner ear stereocilia, immune synapses, endocytic patches, adhesion contacts, and invadosomes of immune and cancer cells. All plastins comprise an N-terminal Ca2 +-binding regulatory headpiece domain followed by two actin-binding domains (ABD1 and ABD2). Actin bundling occurs due to simultaneous binding of both ABDs to separate actin filaments. Bundling is negatively regulated by Ca2 +, but the mechanism of this inhibition remains unknown. In this study, we found that the bundling abilities of PLS1 and PLS2 were similarly sensitive to Ca2 + (pCa50 ~ 6.4), whereas PLS3 was less sensitive (pCa50 ~ 5.9). At the same time, all three isoforms bound to F-actin in a Ca2 +-independent manner, suggesting that binding of only one of the ABDs is inhibited by Ca2 +. Using limited proteolysis and mass spectrometry, we found that in the presence of Ca2 + the EF-hands of human plastins bound to an immediately adjacent sequence homologous to canonical calmodulin-binding peptides. Furthermore, our data from differential centrifugation, Förster resonance energy transfer, native electrophoresis, and chemical crosslinking suggest that Ca2 + does not affect ABD1 but inhibits the ability of ABD2 to interact with actin. A structural mechanism of signal transmission from Ca2 + to ABD2 through EF-hands remains to be established." @default.
• W2732605882 created "2017-07-14" @default.
• W2732605882 creator A5004456531 @default.
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• W2732605882 creator A5044975181 @default.
• W2732605882 creator A5056042395 @default.
• W2732605882 creator A5085075424 @default.
• W2732605882 date "2017-08-01" @default.
• W2732605882 modified "2023-10-05" @default.
• W2732605882 title "The Roles of Actin-Binding Domains 1 and 2 in the Calcium-Dependent Regulation of Actin Filament Bundling by Human Plastins" @default.
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• W2732605882 doi "https://doi.org/10.1016/j.jmb.2017.06.021" @default.
• W2732605882 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/28694070" @default.
• W2732605882 hasPublicationYear "2017" @default.
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