Protective hinge in insulin opens to enable its receptor engagement

John G. Menting, Yanwu Yang, Shu Jin Chan, Nelson B. Phillips, Brian J. Smith, Jonathan Whittaker, Nalinda P. Wickramasinghe, Linda J. Whittaker, Vijay Pandyarajan, Zhu Li Wan, Satya P. Yadav, Julie M. Carroll, Natalie Strokes, Charles Roberts, Faramarz Ismail-Beigi, Wieslawa Milewski, Donald F. Steiner, Virander S. Chauhan, Colin W. Ward, Michael A. Weiss & 1 others Michael C. Lawrence

    Research output: Contribution to journalArticle

    70 Citations (Scopus)

    Abstract

    Insulin provides a classical model of a globular protein, yet how the hormone changes conformation to engage its receptor has long been enigmatic. Interest has focused on the C-terminal B-chain segment, critical for protective self-assembly in β cells and receptor binding at target tissues. Insight may be obtained from truncated "microreceptors" that reconstitute the primary hormone-binding site (α-subunit domains L1 and αCT). We demonstrate that, on microreceptor binding, this segment undergoes concerted hinge-like rotation at its B20-B23 β-turn, coupling reorientation of PheB24 to a 60° rotation of the B25-B28 β-strand away from the hormone core to lie antiparallel to the receptor's L1-β2 sheet. Opening of this hinge enables conserved nonpolar side chains (Ile A2, ValA3, ValB12, PheB24, and PheB25) to engage the receptor. Restraining the hinge by nonstandard mutagenesis preserves native folding but blocks receptor binding, whereas its engineered opening maintains activity at the price of protein instability and nonnative aggregation. Our findings rationalize properties of clinical mutations in the insulin family and provide a previously unidentified foundation for designing therapeutic analogs. We envisage that a switch between free and receptor-bound conformations of insulin evolved as a solution to conflicting structural determinants of biosynthesis and function.

    Original languageEnglish (US)
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume111
    Issue number33
    DOIs
    StatePublished - Aug 19 2014

    Fingerprint

    Hormones
    Insulin
    varespladib methyl
    Mutagenesis
    Proteins
    Binding Sites
    Mutation
    Therapeutics

    Keywords

    • Diabetes mellitus
    • Metabolism
    • Protein structure
    • Receptor tyrosine kinase
    • Signal transduction

    ASJC Scopus subject areas

    • General

    Cite this

    Menting, J. G., Yang, Y., Chan, S. J., Phillips, N. B., Smith, B. J., Whittaker, J., ... Lawrence, M. C. (2014). Protective hinge in insulin opens to enable its receptor engagement. Proceedings of the National Academy of Sciences of the United States of America, 111(33). https://doi.org/10.1073/pnas.1412897111

    Protective hinge in insulin opens to enable its receptor engagement. / Menting, John G.; Yang, Yanwu; Chan, Shu Jin; Phillips, Nelson B.; Smith, Brian J.; Whittaker, Jonathan; Wickramasinghe, Nalinda P.; Whittaker, Linda J.; Pandyarajan, Vijay; Wan, Zhu Li; Yadav, Satya P.; Carroll, Julie M.; Strokes, Natalie; Roberts, Charles; Ismail-Beigi, Faramarz; Milewski, Wieslawa; Steiner, Donald F.; Chauhan, Virander S.; Ward, Colin W.; Weiss, Michael A.; Lawrence, Michael C.

    In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, No. 33, 19.08.2014.

    Research output: Contribution to journalArticle

    Menting, JG, Yang, Y, Chan, SJ, Phillips, NB, Smith, BJ, Whittaker, J, Wickramasinghe, NP, Whittaker, LJ, Pandyarajan, V, Wan, ZL, Yadav, SP, Carroll, JM, Strokes, N, Roberts, C, Ismail-Beigi, F, Milewski, W, Steiner, DF, Chauhan, VS, Ward, CW, Weiss, MA & Lawrence, MC 2014, 'Protective hinge in insulin opens to enable its receptor engagement', Proceedings of the National Academy of Sciences of the United States of America, vol. 111, no. 33. https://doi.org/10.1073/pnas.1412897111
    Menting, John G. ; Yang, Yanwu ; Chan, Shu Jin ; Phillips, Nelson B. ; Smith, Brian J. ; Whittaker, Jonathan ; Wickramasinghe, Nalinda P. ; Whittaker, Linda J. ; Pandyarajan, Vijay ; Wan, Zhu Li ; Yadav, Satya P. ; Carroll, Julie M. ; Strokes, Natalie ; Roberts, Charles ; Ismail-Beigi, Faramarz ; Milewski, Wieslawa ; Steiner, Donald F. ; Chauhan, Virander S. ; Ward, Colin W. ; Weiss, Michael A. ; Lawrence, Michael C. / Protective hinge in insulin opens to enable its receptor engagement. In: Proceedings of the National Academy of Sciences of the United States of America. 2014 ; Vol. 111, No. 33.
    @article{79034a728ace42059babb6578a31dffa,
    title = "Protective hinge in insulin opens to enable its receptor engagement",
    abstract = "Insulin provides a classical model of a globular protein, yet how the hormone changes conformation to engage its receptor has long been enigmatic. Interest has focused on the C-terminal B-chain segment, critical for protective self-assembly in β cells and receptor binding at target tissues. Insight may be obtained from truncated {"}microreceptors{"} that reconstitute the primary hormone-binding site (α-subunit domains L1 and αCT). We demonstrate that, on microreceptor binding, this segment undergoes concerted hinge-like rotation at its B20-B23 β-turn, coupling reorientation of PheB24 to a 60° rotation of the B25-B28 β-strand away from the hormone core to lie antiparallel to the receptor's L1-β2 sheet. Opening of this hinge enables conserved nonpolar side chains (Ile A2, ValA3, ValB12, PheB24, and PheB25) to engage the receptor. Restraining the hinge by nonstandard mutagenesis preserves native folding but blocks receptor binding, whereas its engineered opening maintains activity at the price of protein instability and nonnative aggregation. Our findings rationalize properties of clinical mutations in the insulin family and provide a previously unidentified foundation for designing therapeutic analogs. We envisage that a switch between free and receptor-bound conformations of insulin evolved as a solution to conflicting structural determinants of biosynthesis and function.",
    keywords = "Diabetes mellitus, Metabolism, Protein structure, Receptor tyrosine kinase, Signal transduction",
    author = "Menting, {John G.} and Yanwu Yang and Chan, {Shu Jin} and Phillips, {Nelson B.} and Smith, {Brian J.} and Jonathan Whittaker and Wickramasinghe, {Nalinda P.} and Whittaker, {Linda J.} and Vijay Pandyarajan and Wan, {Zhu Li} and Yadav, {Satya P.} and Carroll, {Julie M.} and Natalie Strokes and Charles Roberts and Faramarz Ismail-Beigi and Wieslawa Milewski and Steiner, {Donald F.} and Chauhan, {Virander S.} and Ward, {Colin W.} and Weiss, {Michael A.} and Lawrence, {Michael C.}",
    year = "2014",
    month = "8",
    day = "19",
    doi = "10.1073/pnas.1412897111",
    language = "English (US)",
    volume = "111",
    journal = "Proceedings of the National Academy of Sciences of the United States of America",
    issn = "0027-8424",
    number = "33",

    }

    TY - JOUR

    T1 - Protective hinge in insulin opens to enable its receptor engagement

    AU - Menting, John G.

    AU - Yang, Yanwu

    AU - Chan, Shu Jin

    AU - Phillips, Nelson B.

    AU - Smith, Brian J.

    AU - Whittaker, Jonathan

    AU - Wickramasinghe, Nalinda P.

    AU - Whittaker, Linda J.

    AU - Pandyarajan, Vijay

    AU - Wan, Zhu Li

    AU - Yadav, Satya P.

    AU - Carroll, Julie M.

    AU - Strokes, Natalie

    AU - Roberts, Charles

    AU - Ismail-Beigi, Faramarz

    AU - Milewski, Wieslawa

    AU - Steiner, Donald F.

    AU - Chauhan, Virander S.

    AU - Ward, Colin W.

    AU - Weiss, Michael A.

    AU - Lawrence, Michael C.

    PY - 2014/8/19

    Y1 - 2014/8/19

    N2 - Insulin provides a classical model of a globular protein, yet how the hormone changes conformation to engage its receptor has long been enigmatic. Interest has focused on the C-terminal B-chain segment, critical for protective self-assembly in β cells and receptor binding at target tissues. Insight may be obtained from truncated "microreceptors" that reconstitute the primary hormone-binding site (α-subunit domains L1 and αCT). We demonstrate that, on microreceptor binding, this segment undergoes concerted hinge-like rotation at its B20-B23 β-turn, coupling reorientation of PheB24 to a 60° rotation of the B25-B28 β-strand away from the hormone core to lie antiparallel to the receptor's L1-β2 sheet. Opening of this hinge enables conserved nonpolar side chains (Ile A2, ValA3, ValB12, PheB24, and PheB25) to engage the receptor. Restraining the hinge by nonstandard mutagenesis preserves native folding but blocks receptor binding, whereas its engineered opening maintains activity at the price of protein instability and nonnative aggregation. Our findings rationalize properties of clinical mutations in the insulin family and provide a previously unidentified foundation for designing therapeutic analogs. We envisage that a switch between free and receptor-bound conformations of insulin evolved as a solution to conflicting structural determinants of biosynthesis and function.

    AB - Insulin provides a classical model of a globular protein, yet how the hormone changes conformation to engage its receptor has long been enigmatic. Interest has focused on the C-terminal B-chain segment, critical for protective self-assembly in β cells and receptor binding at target tissues. Insight may be obtained from truncated "microreceptors" that reconstitute the primary hormone-binding site (α-subunit domains L1 and αCT). We demonstrate that, on microreceptor binding, this segment undergoes concerted hinge-like rotation at its B20-B23 β-turn, coupling reorientation of PheB24 to a 60° rotation of the B25-B28 β-strand away from the hormone core to lie antiparallel to the receptor's L1-β2 sheet. Opening of this hinge enables conserved nonpolar side chains (Ile A2, ValA3, ValB12, PheB24, and PheB25) to engage the receptor. Restraining the hinge by nonstandard mutagenesis preserves native folding but blocks receptor binding, whereas its engineered opening maintains activity at the price of protein instability and nonnative aggregation. Our findings rationalize properties of clinical mutations in the insulin family and provide a previously unidentified foundation for designing therapeutic analogs. We envisage that a switch between free and receptor-bound conformations of insulin evolved as a solution to conflicting structural determinants of biosynthesis and function.

    KW - Diabetes mellitus

    KW - Metabolism

    KW - Protein structure

    KW - Receptor tyrosine kinase

    KW - Signal transduction

    UR - http://www.scopus.com/inward/record.url?scp=84906306629&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84906306629&partnerID=8YFLogxK

    U2 - 10.1073/pnas.1412897111

    DO - 10.1073/pnas.1412897111

    M3 - Article

    VL - 111

    JO - Proceedings of the National Academy of Sciences of the United States of America

    JF - Proceedings of the National Academy of Sciences of the United States of America

    SN - 0027-8424

    IS - 33

    ER -