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Basic amino-acid side chains regulate transmembrane integrin signalling.

ABSTRACT: Side chains of Lys/Arg near transmembrane domain (TMD) membrane-water interfaces can 'snorkel', placing their positive charge near negatively charged phospholipid head groups; however, snorkelling's functional effects are obscure. Integrin ? TMDs have such conserved basic amino acids. Here we use NMR spectroscopy to show that integrin ?(3)(Lys?716) helps determine ?(3) TMD topography. The ?(??b)?(3) TMD structure indicates that precise ?(3) TMD crossing angles enable the assembly of outer and inner membrane 'clasps' that hold the ?? TMD together to limit transmembrane signalling. Mutation of ?(3)(Lys?716) caused dissociation of ?(??b)?(3) TMDs and integrin activation. To confirm that altered topography of ?(3)(Lys?716) mutants activated ?(??b)?(3), we used directed evolution of ?(3)(K716A) to identify substitutions restoring default state. Introduction of Pro(711) at the midpoint of ?(3) TMD (A711P) increased ?(??b)?(3) TMD association and inactivated integrin ?(??b)?(3)(A711P,K716A). ?(3)(Pro?711) introduced a TMD kink of 30?±?1° precisely at the border of the outer and inner membrane clasps, thereby decoupling the tilt between these segments. Thus, widely occurring snorkelling residues in TMDs can help maintain TMD topography and membrane-embedding, thereby regulating transmembrane signalling.

SUBMITTER: Kim C

PROVIDER: S-EPMC3257387 | biostudies-literature | 2011 Dec

REPOSITORIES: biostudies-literature

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Basic amino-acid side chains regulate transmembrane integrin signalling.

Kim Chungho C Schmidt Thomas T Cho Eun-Gyung EG Ye Feng F Ulmer Tobias S TS Ginsberg Mark H MH

Nature 20111218 7380

Side chains of Lys/Arg near transmembrane domain (TMD) membrane-water interfaces can 'snorkel', placing their positive charge near negatively charged phospholipid head groups; however, snorkelling's functional effects are obscure. Integrin β TMDs have such conserved basic amino acids. Here we use NMR spectroscopy to show that integrin β(3)(Lys 716) helps determine β(3) TMD topography. The α(ΙΙb)β(3) TMD structure indicates that precise β(3) TMD crossing angles enable the assembly of outer and in ...[more]

PMID: 22178926

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Project description:Amino acid side-chain conformational properties influence the overall structural and dynamic properties of proteins and, therefore, their biological functions. In this study, quantum mechanical (QM) potential energy surfaces for the rotation of side-chain χ(1) and χ(2) torsions in dipeptides in the alphaR, beta, and alphaL backbone conformations were calculated. The QM energy surfaces provide a broad view of the intrinsic conformational properties of each amino acid side-chain. The extent to which intrinsic energetics dictates side-chain orientation was studied through comparisons of the QM energy surfaces with χ(1) and χ(2) free energy surfaces from probability distributions obtained from a survey of high resolution crystal structures. In general, the survey probability maxima are centered in minima of the QM surfaces as expected for sp(3) (or sp(2) for χ(2) of Asn, Phe, Trp, and Tyr) atom centers with strong variations between amino acids occurring in the energies of the minima indicating intrinsic differences in rotamer preferences. High correlations between the QM and survey data were found for hydrophobic side-chains except Met, suggesting minimal influence of the protein and solution environments on their conformational distributions. Conversely, low correlations for polar or charged side-chains indicate a dominant role of the environment in stabilizing conformations that are not intrinsically favored. Data also link the presence of off-rotamers in His and Trp to favorable interactions with the backbone. Results also suggest that the intrinsic energetics of the side-chains of Phe and Tyr may play important roles in protein folding and stability. Analyses on whether intrinsic side-chain energetics can influence backbone preference identified a strong correlation for residues in the alphaL backbone conformation. It is suggested that this correlation reflects the intrinsic instability of the alphaL backbone such that assumption of this backbone conformation is facilitated by intrinsically favorable side-chain conformations. Together our results offer a broad overview of the conformational properties of amino acid side-chains and the QM data may be used as target data for force field optimization.

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Basic amino-acid side chains regulate transmembrane integrin signalling.

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Basic amino-acid side chains regulate transmembrane integrin signalling.

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