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Collagen-binding mode of vWF-A3 domain determined by a transferred cross-saturation experiment

Nature Structural Biology volume 10, pages 53–58 (2003)Cite this article

Abstract

The nature of the supramolecular complex between fibrillar collagen and collagen-binding proteins (CBPs) has hindered detailed X-ray and NMR analyses of the ligand-recognition mechanism at atomic resolution because of the lack of appropriate approaches for studying large heterogeneous supramolecular complexes. Recently, we proposed an NMR method, termed transferred cross-saturation (TCS), that enables the rigorous identification of contact residues in a huge protein complex. Here we used TCS to study the supramolecular complex between the A3 domain of von Willebrand factor and fibrillar collagen, which allowed the successful determination of the ligand-binding site of the A3 domain. The binding site of the A3 domain was located at its hydrophobic 'front' surface and was completely different from that of the I domain from the a2 subunit of integrin (α2-I domain), which was reported to be the hydrophilic 'top' surface of α2-I, although the A3 domain and the α2-I domain share a similar fold and possess the identical function of collagen binding.

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Figure 1: Overview and results of TCS experiments.
Figure 2: Mapping of the affected residues in the TCS experiments on the A3 domain and the results of the mutational experiments.
Figure 3: Surface representation of the binding trenches of the A3 domain.
Figure 4: Comparison of the collagen-binding sites between the A3 domain and the α2-I domain.

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Acknowledgements

I.S. thanks G. Wagner and M. Ikura for useful discussions. This work was supported by a grant from the Japan New Energy and Industrial Technology Development Organization (NEDO).

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Authors and Affiliations

  1. Graduate School of Pharmaceutical Sciences, the University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Noritaka Nishida, Masayoshi Sakakura, Hiroaki Terasawa & Ichio Shimada

  2. Japan Biological Information Research Center (JBIRC), Japan Biological Informatics Consortium (JBIC), Hatchobori, Chuo-ku, Tokyo, 104-0032, Japan

    Noritaka Nishida, Hiromi Sumikawa, Nobuhisa Shimba & Ei-ichiro Suzuki

  3. Institute of Life Sciences, Ajinomoto Co., Inc., Suzuki-cho, Kawasaki-ku, Kawasaki, 210-8681, Japan

    Hiromi Sumikawa, Nobuhisa Shimba & Ei-ichiro Suzuki

  4. Biological Information Research Center (BIRC), National Institute of Advanced Industrial Science and Technology (AIST), Aomi, Koto-ku, Tokyo, 135-0064, Japan

    Hideo Takahashi & Ichio Shimada

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  1. Noritaka Nishida
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Correspondence to Ichio Shimada.

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Nishida, N., Sumikawa, H., Sakakura, M. et al. Collagen-binding mode of vWF-A3 domain determined by a transferred cross-saturation experiment. Nat Struct Mol Biol 10, 53–58 (2003). https://doi.org/10.1038/nsb876

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