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Basic amino-acid side chains regulate transmembrane integrin signalling

Nature volume 481, pages 209–213 (2012)Cite this article

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An Erratum to this article was published on 08 May 2013

Abstract

Side chains of Lys/Arg near transmembrane domain (TMD)1,2,3 membrane–water interfaces can ‘snorkel’, placing their positive charge near negatively charged phospholipid head groups4,5,6; however, snorkelling’s functional effects are obscure. Integrin β TMDs have such conserved basic amino acids. Here we use NMR spectroscopy7,8 to show that integrin β3(Lys 716) helps determine β3 TMD topography. The αΙΙbβ3 TMD structure indicates that precise β3 TMD crossing angles enable the assembly of outer and inner membrane ‘clasps’ that hold the αβ TMD together to limit transmembrane signalling9. Mutation of β3(Lys 716) caused dissociation of αΙΙbβ3 TMDs and integrin activation. To confirm that altered topography of β3(Lys 716) mutants activated αΙΙbβ3, we used directed evolution of β3(K716A) to identify substitutions restoring default state. Introduction of Pro(711) at the midpoint of β3 TMD (A711P) increased αΙΙbβ3 TMD association and inactivated integrin αΙΙbβ3(A711P,K716A). β3(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.

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Figure 1: Loss of snorkelling lysine changes lipid embedding of β 3 TMD.
Figure 2: Mutations in the snorkelling lysine induce integrin activation and disrupt α-β TMD interaction.
Figure 3: Directed evolution of the β 3 integrin to identfy mutations that complement the activating effect of snorkelling lysine mutation.
Figure 4: Proline introduced in the TMD forms a flexible kink that stabilizes the α ΙΙb β 3 (K716A) TMD interaction and reduces integrin activation.

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Acknowledgements

This work was supported by grants from the National Institutes of Health of the USA. T.S.U. acknowledges support from the National Institutes of Health (HL089726) and M.H.G. was supported by HL078784, HL57900 and AR27214. C.K. is a recipient of a postdoctoral fellowship from the American Institute for Cancer Research.

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Author notes

  1. Chungho Kim and Thomas Schmidt: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medicine, University of California, San Diego, La Jolla, California 92093, USA,

    Chungho Kim, Feng Ye & Mark H. Ginsberg

  2. Department of Biochemistry and Molecular Biology, Zilkha Neurogenetic Institute, Keck School of Medicine, University of Southern California, Los Angeles, 90033, California, USA

    Thomas Schmidt & Tobias S. Ulmer

  3. Center for Neuroscience, Aging, and Stem Cell Research, Sanford-Burnham Medical Research Institute, La Jolla, California 92037, USA,

    Eun-Gyung Cho

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  1. Chungho Kim
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  6. Mark H. Ginsberg
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Contributions

The project was conceived by C.K. and M.H.G. All experiments with the exception of the NMR studies were performed by C.K. The NMR studies were conducted by T.S. under the supervision of T.S.U. E.C. and F.Y. contributed reagents. M.H.G. and C.K. wrote the paper, which was edited by T.S. and T.S.U.

Corresponding authors

Correspondence to Tobias S. Ulmer or Mark H. Ginsberg.

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The authors declare no competing financial interests.

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Kim, C., Schmidt, T., Cho, EG. et al. Basic amino-acid side chains regulate transmembrane integrin signalling. Nature 481, 209–213 (2012). https://doi.org/10.1038/nature10697

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