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A phenylalanine zipper mediates APS dimerization

Nature Structural & Molecular Biology volume 11, pages 968–974 (2004)Cite this article

Abstract

The APS, SH2-B and LNK proteins are adapters that activate and modulate receptor tyrosine kinase and JAK/STAT signaling. We now show that a conserved N-terminal domain mediates APS homodimerization. We determined the crystal structure of the dimerization domain at a resolution of 1.7 Å using bromide ion MAD phasing. Each molecule contributes two helices to a compact four-helix bundle having a bisecting-U topology. Its most conspicuous feature is a stack of interdigitated phenylalanine side chains at the domain core. These residues create a new motif we refer to as a 'phenylalanine zipper,' which is critical to dimerization. A newly developed bridging yeast tri-hybrid assay showed that APS dimerizes JAK2, insulin receptor and IGF1 receptor kinases using its SH2 and dimerization domains. Dimerization via the phenylalanine zipper domain provides a mechanism for activating and modulating tyrosine kinase activity even in the absence of extracellular ligands.

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Figure 1: The APS/SH2-B/Lnk family.
Figure 2: Yeast two-hybrid assays and analytical ultracentrifugation.
Figure 3: Crystal structure of the APS dimerization domain.
Figure 4: Intersection of αN and αC′.
Figure 5: Functional significance of APS dimerization.

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Acknowledgements

We thank A. Saxena and the staff of beamline X12C at National Synchrotron Light Source and D. Frantz and J. Lee for helpful discussions and assistance. These studies were funded by US National Institutes of Health (NIH) grant R01 DK43123 (S.E.S.), Joslin Diabetes and Endocrinology Research Center NIH grant DK36836, fellowships from the Mary K. Iacocca Foundation (S.D. and Y.-I.C.), NIH (R29 DK09393 to E.D.W.), and Sankyo Foundation (M.N.), and the Helen and Morton Adler Chair in Structural Biology (S.E.S.).

Author information

Author notes

  1. Sirano Dhe-Paganon

    Present address: Department of Physiology, University of Toronto, Toronto, Ontario, Canada

  2. Masahiro Nishi

    Present address: The First Department of Medicine, Wakayama University of Medical Science, Wakayama, Japan

  3. Young-In Chi

    Present address: Structural Biology Center, Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, 40536, USA

  4. Sirano Dhe-Paganon, Eric D Werner and Masahiro Nishi: These authors contributed equally to this work.

Authors and Affiliations

  1. Joslin Diabetes Center and Department of Medicine, Harvard Medical School, Boston, 02215, Massachusetts, USA

    Sirano Dhe-Paganon, Eric D Werner, Masahiro Nishi, Lone Hansen, Young-In Chi & Steven E Shoelson

  2. Department of Physiology, University of Toronto, Toronto, Ontario, Canada

    Masahiro Nishi & Young-In Chi

  3. The First Department of Medicine, Wakayama University of Medical Science, Wakayama, Japan

    Sirano Dhe-Paganon & Young-In Chi

  4. Structural Biology Center, Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, 40536, USA

    Sirano Dhe-Paganon & Masahiro Nishi

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  1. Sirano Dhe-Paganon
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Correspondence to Steven E Shoelson.

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Supplementary information

Supplementary Fig. 1

Electron density and controls. (PDF 168 kb)

Supplementary Table 1

MAD phasing data. (PDF 17 kb)

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Dhe-Paganon, S., Werner, E., Nishi, M. et al. A phenylalanine zipper mediates APS dimerization. Nat Struct Mol Biol 11, 968–974 (2004). https://doi.org/10.1038/nsmb829

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