5l81

Publication Abstract from PubMed

Kindlins co-activate integrins alongside talin. They possess, like talin, a FERM domain comprising F0-F3 subdomains, but with a pleckstrin homology (PH) domain inserted in the F2 subdomain that enables membrane association. We present the crystal structure of murine kindlin-3 PH domain determined at 2.23A resolution and characterise its lipid binding using biophysical and computational approaches. Molecular dynamics (MD) simulations suggest flexibility in the PH domain loops connecting beta-strands forming the putative phosphatidylinositol phosphate (PtdInsP) binding site. Simulations with PtdInsP-containing bilayers reveal that the PH domain associates with PtdInsP molecules mainly via the positively charged surface presented by the beta1-beta2 loop and that it binds with somewhat higher affinity to PtdIns(3,4,5)P3 compared to PtdIns(4,5)P2 Surface plasmon resonance (SPR) with lipid headgroups immobilised and the PH domain as analyte indicate affinities of 300 muM for PtdIns(3,4,5)P3 and 1mM for PtdIns(4,5)P2 In contrast, SPR studies with immobilised PH domain and lipid nanodiscs as analyte show affinities of 0.40 microM for PtdIns(3,4,5)P3 and no affinity for PtdIns(4,5)P2 when the inositol phosphate constitutes 5% of the total lipids (~5 molecules per nanodisc). Reducing the PtdIns(3,4,5)P3 composition to 1% abolishes nanodisc binding to the PH domain, as does site-directed mutagenesis of two lysines within the beta1-beta2 loop. Binding of PtdIns(3,4,5)P3 by a canonical PH domain, Grp1, is not similarly influenced by SPR experimental design. These data suggest a role for PtdIns(3,4,5)P3 clustering in the binding of some PH domains and not others, highlighting the importance lipid mobility and clustering for the biophysical assessment of protein-membrane interactions.

Structure and lipid binding properties of the kindlin-3 pleckstrin homology domain.,Ni T, Kalli A, Naughton F, Yates L, Naneh O, Kozorog M, Anderluh G, Sansom M, Gilbert R Biochem J. 2016 Dec 14. pii: BCJ20160791. PMID:27974389^[4]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.