The biochemical effect of Ser167 phosphorylation on Chlamydomonas reinhardtii centrin

Susan M. Meyn, Christina Seda, Muriel Campbell, Kevin L. Weiss, Haitao Hu, Belinda Pastrana-Rios, Walter J. Chazin

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Centrin is an EF-hand calcium-binding protein found in microtubule organizing centers of organisms ranging from algae and yeast to man. Phosphorylation in the centrin C-terminal domain occurs in mitosis and is associated with alterations in contractile fibers. To obtain insight into the structural basis for the functional effect of phosphorylation, Chlamydomonas reinhardtii centrin C-terminal domain phosphorylated at Ser167 (pCRC-C) has been produced and characterized. The structure of pCRC-C was compared to the unmodified protein by NMR spectroscopy. The effect of phosphorylation on target binding was examined for the complex of pCRC-C and a 19 residue centrin-binding fragment of Kar1. Remarkably, the efficient and selective phosphorylation by PKA was suppressed in the complex. Moreover, comparisons of NMR chemical shift differences induced by phosphorylation reveal a greater effect from phosphorylation in the context of the Kar1 complex than for the free protein. These results directly demonstrate that phosphorylation modulates the structure and biochemical activities of centrin.

Original languageEnglish
Pages (from-to)342-348
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume342
Issue number1
DOIs
StatePublished - Mar 31 2006

Funding

This work was supported by the National Institutes of Health Grants RO1 GM-40120 and RO1 GM 65484 (to W.J.C.), NIH-SCORE Grant 5-S06GM08103 (to B.P.-R.), and R25 GM67072 (to Vanderbilt Summer Research Experiences for Undergraduates), with facilities supported by the Vanderbilt Center in Molecular Toxicology Grant (P50 ES00267) and Vanderbilt Ingram Cancer Center (P30 CA68485).

FundersFunder number
NIH-SCORER25 GM67072, 5-S06GM08103
Vanderbilt Center in Molecular ToxicologyP50 ES00267
National Institutes of HealthRO1 GM 65484
National Institute of General Medical SciencesR01GM040120
Vanderbilt-Ingram Cancer CenterP30 CA68485

    Keywords

    • Calcium sensor
    • Calmodulin
    • Caltractin
    • Centrin
    • Conformational exchange
    • EF-hand calcium-binding protein
    • Energetic coupling
    • Kar1

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