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Sequential assignment of proline-rich regions in proteins: Application to modular binding domain complexes

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Abstract

Many protein–protein interactions involve amino acid sequences containing proline-rich motifs and even poly-proline stretches. The lack of amide protons in such regions complicates assignment, since 1HN-based triple-resonance assignment strategies cannot be employed. Two such systems that we are currently studying include an SH2 domain from the protein Crk with a region containing 9 prolines in a 14 amino acid sequence, as well as a WW domain that interacts with a proline-rich target. A modified version of the HACAN pulse scheme, originally described by Bax and co-workers [Wang et al. (1995) J. Biomol. NMR, 5, 376–382], and an experiment which correlates the intra-residue 1Hα, 13Cα/13Cβ chemical shifts with the 15N shift of the subsequent residue are presented and applied to the two systems listed above, allowing sequential assignment of the molecules.

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Authors and Affiliations

  1. Department of Biochemistry, The University of Toronto, Canada

    Voula Kanelis, Daniela Rotin & Julie D. Forman-Kay

  2. Program in Structural Biology and Biochemistry, The Hospital for Sick Children, Toronto, ON, Canada, G 1X8

    Voula Kanelis & Julie D. Forman-Kay

  3. Protein Engineering Network Centres of Excellence and Departments of Medical Genetics, Biochemistry and Chemistry, The University of Toronto, Toronto, ON, Canada, M5S 1A8

    Logan Donaldson, D.R. Muhandiram & Lewis E. Kay

  4. Program in Cell Biology, The Hospital for Sick Children, Toronto, ON, Canada, M5G 1X8

    Daniela Rotin

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  1. Voula Kanelis
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  2. Logan Donaldson
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  3. D.R. Muhandiram
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  4. Daniela Rotin
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  5. Julie D. Forman-Kay
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  6. Lewis E. Kay
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Kanelis, V., Donaldson, L., Muhandiram, D. et al. Sequential assignment of proline-rich regions in proteins: Application to modular binding domain complexes. J Biomol NMR 16, 253–259 (2000). https://doi.org/10.1023/A:1008355012528

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