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Structural and functional insights into PINCH LIM4 domain–mediated integrin signaling

Nature Structural & Molecular Biology volume 10, pages 558–564 (2003)Cite this article

Abstract

PINCH is an adaptor protein found in focal adhesions, large cellular complexes that link extracellular matrix to the actin cytoskeleton. PINCH, which contains an array of five LIM domains, has been implicated as a platform for multiple protein–protein interactions that mediate integrin signaling within focal adhesions. We had previously characterized the LIM1 domain of PINCH, which functions in focal adhesions by binding specifically to integrin-linked kinase. Using NMR spectroscopy, we show here that the PINCH LIM4 domain, while maintaining the conserved LIM scaffold, recognizes the third SH3 domain of another adaptor protein, Nck2 (also called Nckβ or Grb4), in a manner distinct from that of the LIM1 domain. Point mutation of LIM residues in the SH3-binding interface disrupted LIM–SH3 interaction and substantially impaired localization of PINCH to focal adhesions. These data provide novel structural insight into LIM domain–mediated protein–protein recognition and demonstrate that the PINCH-Nck2 interaction is an important component of the focal adhesion assembly during integrin signaling.

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Figure 1: Structural properties of LIM4 and its sequence and structural comparisons with other LIM domains.
Figure 2: Chemical shift perturbation data for the LIM4–SH3-3 interaction.
Figure 3: ILK-PINCH and PINCH-Nck2 interactions.
Figure 4: R197A-R198A double mutation reduces the efficiency of PINCH localization to focal adhesions.

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Acknowledgements

We thank F. Delaglio for NMRPipe software and D. Garrett for PIPP. The work was supported by US National Institutes of Health grants to J.Q. and C.W.

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Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, Case Western Reserve University, 10500 Euclid Avenue, Cleveland, 44102, Ohio, USA

    Algirdas Velyvis & Jun Qin

  2. Structural Biology Program, Lerner Research Institute, The Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, 44195, Ohio, USA

    Algirdas Velyvis, Julia Vaynberg, Yanwu Yang, Olga Vinogradova & Jun Qin

  3. Department of Pathology, University of Pittsburgh, 707B Scaife Hall, 3550 Terrace Street, Pittsburgh, 15261, Pennsylvania, USA

    Yongjun Zhang & Chuanyue Wu

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  1. Algirdas Velyvis
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Correspondence to Chuanyue Wu or Jun Qin.

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Velyvis, A., Vaynberg, J., Yang, Y. et al. Structural and functional insights into PINCH LIM4 domain–mediated integrin signaling. Nat Struct Mol Biol 10, 558–564 (2003). https://doi.org/10.1038/nsb938

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