How insulin binds: The B-chain α-helix contacts the L1 β-helix of the insulin receptor

Kun Huang, Bin Xu, Shi Quan Hu, Ying Chi Chu, Qing Xin Hua, Yan Qu, Biaoru Li, Shuhua Wang, Run Ying Wang, Satoe H. Nakagawa, Anne Mette Theede, Jonathan Whittaker, Pierre De Meyts, Panayotis G. Katsoyannis, Michael A. Weiss

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Binding of insulin to the insulin receptor plays a central role in the hormonal control of metabolism. Here, we investigate possible contact sites between the receptor and the conserved non-polar surface of the B-chain. Evidence is presented that two contiguous sites in an α-helix, Val B12 and TyrB16, contact the receptor. Chemical synthesis is exploited to obtain non-standard substitutions in an engineered monomer (DKP-insulin). Substitution of TyrB16 by an isosteric photo-activatable derivative (para-azido-phenylalanine) enables efficient cross-linking to the receptor. Such cross-linking is specific and maps to the L1 β-helix of the α-subunit. Because substitution of ValB12 by larger side-chains markedly impairs receptor binding, cross-linking studies at B12 were not undertaken. Structure-function relationships are instead probed by side-chains of similar or smaller volume: respective substitution of Val B12 by alanine, threonine, and α-aminobutyric acid leads to activities of 1(±0.1)%, 13(±6)%, and 14(±5)% (relative to DKP-insulin) without disproportionate changes in negative cooperativity. NMR structures are essentially identical with native insulin. The absence of transmitted structural changes suggests that the low activities of B12 analogues reflect local perturbation of a "high-affinity" hormone-receptor contact. By contrast, because position B16 tolerates alanine substitution (relative activity 34(±10)%), the contribution of this neighboring interaction is smaller. Together, our results support a model in which the B-chain α-helix, functioning as an essential recognition element, docks against the L1 β-helix of the insulin receptor.

Original languageEnglish (US)
Pages (from-to)529-550
Number of pages22
JournalJournal of Molecular Biology
Volume341
Issue number2
DOIs
StatePublished - Aug 6 2004

Fingerprint

Insulin Receptor
Alanine
Insulin
Aminobutyrates
Threonine
Phenylalanine
Hormones
DKP-insulin

Keywords

  • Aba, α-aminobutyric acid
  • CD, circular dichroism
  • CHO, Chinese hamster ovary
  • CR, cysteine-rich domain of receptor α-subunit
  • DG, distance geometry
  • NMR spectroscopy
  • diabetes mellitus
  • metabolism
  • non-standard mutagenesis
  • polypeptide hormone

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Huang, K., Xu, B., Hu, S. Q., Chu, Y. C., Hua, Q. X., Qu, Y., ... Weiss, M. A. (2004). How insulin binds: The B-chain α-helix contacts the L1 β-helix of the insulin receptor. Journal of Molecular Biology, 341(2), 529-550. https://doi.org/10.1016/j.jmb.2004.05.023

How insulin binds : The B-chain α-helix contacts the L1 β-helix of the insulin receptor. / Huang, Kun; Xu, Bin; Hu, Shi Quan; Chu, Ying Chi; Hua, Qing Xin; Qu, Yan; Li, Biaoru; Wang, Shuhua; Wang, Run Ying; Nakagawa, Satoe H.; Theede, Anne Mette; Whittaker, Jonathan; De Meyts, Pierre; Katsoyannis, Panayotis G.; Weiss, Michael A.

In: Journal of Molecular Biology, Vol. 341, No. 2, 06.08.2004, p. 529-550.

Research output: Contribution to journalArticle

Huang, K, Xu, B, Hu, SQ, Chu, YC, Hua, QX, Qu, Y, Li, B, Wang, S, Wang, RY, Nakagawa, SH, Theede, AM, Whittaker, J, De Meyts, P, Katsoyannis, PG & Weiss, MA 2004, 'How insulin binds: The B-chain α-helix contacts the L1 β-helix of the insulin receptor', Journal of Molecular Biology, vol. 341, no. 2, pp. 529-550. https://doi.org/10.1016/j.jmb.2004.05.023
Huang, Kun ; Xu, Bin ; Hu, Shi Quan ; Chu, Ying Chi ; Hua, Qing Xin ; Qu, Yan ; Li, Biaoru ; Wang, Shuhua ; Wang, Run Ying ; Nakagawa, Satoe H. ; Theede, Anne Mette ; Whittaker, Jonathan ; De Meyts, Pierre ; Katsoyannis, Panayotis G. ; Weiss, Michael A. / How insulin binds : The B-chain α-helix contacts the L1 β-helix of the insulin receptor. In: Journal of Molecular Biology. 2004 ; Vol. 341, No. 2. pp. 529-550.
@article{c210058c0c2c4ceab31655f6aeb2952b,
title = "How insulin binds: The B-chain α-helix contacts the L1 β-helix of the insulin receptor",
abstract = "Binding of insulin to the insulin receptor plays a central role in the hormonal control of metabolism. Here, we investigate possible contact sites between the receptor and the conserved non-polar surface of the B-chain. Evidence is presented that two contiguous sites in an α-helix, Val B12 and TyrB16, contact the receptor. Chemical synthesis is exploited to obtain non-standard substitutions in an engineered monomer (DKP-insulin). Substitution of TyrB16 by an isosteric photo-activatable derivative (para-azido-phenylalanine) enables efficient cross-linking to the receptor. Such cross-linking is specific and maps to the L1 β-helix of the α-subunit. Because substitution of ValB12 by larger side-chains markedly impairs receptor binding, cross-linking studies at B12 were not undertaken. Structure-function relationships are instead probed by side-chains of similar or smaller volume: respective substitution of Val B12 by alanine, threonine, and α-aminobutyric acid leads to activities of 1(±0.1){\%}, 13(±6){\%}, and 14(±5){\%} (relative to DKP-insulin) without disproportionate changes in negative cooperativity. NMR structures are essentially identical with native insulin. The absence of transmitted structural changes suggests that the low activities of B12 analogues reflect local perturbation of a {"}high-affinity{"} hormone-receptor contact. By contrast, because position B16 tolerates alanine substitution (relative activity 34(±10){\%}), the contribution of this neighboring interaction is smaller. Together, our results support a model in which the B-chain α-helix, functioning as an essential recognition element, docks against the L1 β-helix of the insulin receptor.",
keywords = "Aba, α-aminobutyric acid, CD, circular dichroism, CHO, Chinese hamster ovary, CR, cysteine-rich domain of receptor α-subunit, DG, distance geometry, NMR spectroscopy, diabetes mellitus, metabolism, non-standard mutagenesis, polypeptide hormone",
author = "Kun Huang and Bin Xu and Hu, {Shi Quan} and Chu, {Ying Chi} and Hua, {Qing Xin} and Yan Qu and Biaoru Li and Shuhua Wang and Wang, {Run Ying} and Nakagawa, {Satoe H.} and Theede, {Anne Mette} and Jonathan Whittaker and {De Meyts}, Pierre and Katsoyannis, {Panayotis G.} and Weiss, {Michael A.}",
year = "2004",
month = "8",
day = "6",
doi = "10.1016/j.jmb.2004.05.023",
language = "English (US)",
volume = "341",
pages = "529--550",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - How insulin binds

T2 - The B-chain α-helix contacts the L1 β-helix of the insulin receptor

AU - Huang, Kun

AU - Xu, Bin

AU - Hu, Shi Quan

AU - Chu, Ying Chi

AU - Hua, Qing Xin

AU - Qu, Yan

AU - Li, Biaoru

AU - Wang, Shuhua

AU - Wang, Run Ying

AU - Nakagawa, Satoe H.

AU - Theede, Anne Mette

AU - Whittaker, Jonathan

AU - De Meyts, Pierre

AU - Katsoyannis, Panayotis G.

AU - Weiss, Michael A.

PY - 2004/8/6

Y1 - 2004/8/6

N2 - Binding of insulin to the insulin receptor plays a central role in the hormonal control of metabolism. Here, we investigate possible contact sites between the receptor and the conserved non-polar surface of the B-chain. Evidence is presented that two contiguous sites in an α-helix, Val B12 and TyrB16, contact the receptor. Chemical synthesis is exploited to obtain non-standard substitutions in an engineered monomer (DKP-insulin). Substitution of TyrB16 by an isosteric photo-activatable derivative (para-azido-phenylalanine) enables efficient cross-linking to the receptor. Such cross-linking is specific and maps to the L1 β-helix of the α-subunit. Because substitution of ValB12 by larger side-chains markedly impairs receptor binding, cross-linking studies at B12 were not undertaken. Structure-function relationships are instead probed by side-chains of similar or smaller volume: respective substitution of Val B12 by alanine, threonine, and α-aminobutyric acid leads to activities of 1(±0.1)%, 13(±6)%, and 14(±5)% (relative to DKP-insulin) without disproportionate changes in negative cooperativity. NMR structures are essentially identical with native insulin. The absence of transmitted structural changes suggests that the low activities of B12 analogues reflect local perturbation of a "high-affinity" hormone-receptor contact. By contrast, because position B16 tolerates alanine substitution (relative activity 34(±10)%), the contribution of this neighboring interaction is smaller. Together, our results support a model in which the B-chain α-helix, functioning as an essential recognition element, docks against the L1 β-helix of the insulin receptor.

AB - Binding of insulin to the insulin receptor plays a central role in the hormonal control of metabolism. Here, we investigate possible contact sites between the receptor and the conserved non-polar surface of the B-chain. Evidence is presented that two contiguous sites in an α-helix, Val B12 and TyrB16, contact the receptor. Chemical synthesis is exploited to obtain non-standard substitutions in an engineered monomer (DKP-insulin). Substitution of TyrB16 by an isosteric photo-activatable derivative (para-azido-phenylalanine) enables efficient cross-linking to the receptor. Such cross-linking is specific and maps to the L1 β-helix of the α-subunit. Because substitution of ValB12 by larger side-chains markedly impairs receptor binding, cross-linking studies at B12 were not undertaken. Structure-function relationships are instead probed by side-chains of similar or smaller volume: respective substitution of Val B12 by alanine, threonine, and α-aminobutyric acid leads to activities of 1(±0.1)%, 13(±6)%, and 14(±5)% (relative to DKP-insulin) without disproportionate changes in negative cooperativity. NMR structures are essentially identical with native insulin. The absence of transmitted structural changes suggests that the low activities of B12 analogues reflect local perturbation of a "high-affinity" hormone-receptor contact. By contrast, because position B16 tolerates alanine substitution (relative activity 34(±10)%), the contribution of this neighboring interaction is smaller. Together, our results support a model in which the B-chain α-helix, functioning as an essential recognition element, docks against the L1 β-helix of the insulin receptor.

KW - Aba, α-aminobutyric acid

KW - CD, circular dichroism

KW - CHO, Chinese hamster ovary

KW - CR, cysteine-rich domain of receptor α-subunit

KW - DG, distance geometry

KW - NMR spectroscopy

KW - diabetes mellitus

KW - metabolism

KW - non-standard mutagenesis

KW - polypeptide hormone

UR - http://www.scopus.com/inward/record.url?scp=3342960841&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3342960841&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2004.05.023

DO - 10.1016/j.jmb.2004.05.023

M3 - Article

C2 - 15276842

AN - SCOPUS:3342960841

VL - 341

SP - 529

EP - 550

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 2

ER -