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NMR structure and binding studies confirm that PA4608 from Pseudomonas aeruginosa is a PilZ domain and a c-di-GMP binding protein

Authors

  • Theresa A. Ramelot,

    1. Pacific Northwest National Laboratory, Biological Sciences Division, Richland, Washington 99354
    2. Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056
    3. Northeast Structural Economics Consortium
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  • Adelinda Yee,

    1. Division of Cancer Genomics and Proteomics, Ontario Cancer Institute Toronto, Ontario M5G 2M9, Canada
    2. Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 2M9, Canada
    3. Northeast Structural Economics Consortium
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  • John R. Cort,

    1. Pacific Northwest National Laboratory, Biological Sciences Division, Richland, Washington 99354
    2. Northeast Structural Economics Consortium
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  • Anthony Semesi,

    1. Division of Cancer Genomics and Proteomics, Ontario Cancer Institute Toronto, Ontario M5G 2M9, Canada
    2. Northeast Structural Economics Consortium
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  • Cheryl H. Arrowsmith,

    1. Division of Cancer Genomics and Proteomics, Ontario Cancer Institute Toronto, Ontario M5G 2M9, Canada
    2. Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 2M9, Canada
    3. Northeast Structural Economics Consortium
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  • Michael A. Kennedy

    Corresponding author
    1. Pacific Northwest National Laboratory, Biological Sciences Division, Richland, Washington 99354
    2. Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056
    3. Northeast Structural Economics Consortium
    • Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056
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Abstract

PA4608 is a 125 residue protein from Pseudomonas aeruginosa with a recent identification as a PilZ domain and putative bis-(3′-5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) adaptor protein that plays a role in bacterial second-messenger regulated processes. The nuclear magnetic resonance (NMR) structure of PA4608 has been determined and c-di-GMP binding has been confirmed by NMR titration studies. The monomeric core structure of PA4608 contains a six-stranded anti-parallel β barrel flanked by three helices. Conserved surface residues among PA4608 homologs suggest the c-di-GMP binding site is at one end of the barrel and includes residues in the helices as well as in the unstructured N-terminus. Chemical shift changes in PA4608 resonances upon titration with c-di-GMP confirm binding. This evidence supports the hypothesis that proteins containing PilZ domains are the long-sought c-di-GMP adaptor proteins. Proteins 2007. © 2006 Wiley-Liss, Inc.

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