Identification of inositol 1,4,5-trisphosphate-binding proteins by heparin-agarose affinity purification and LTQ ORBITRAP MS in Oryza sativa

Authors

  • Yanli Nie,

    1. Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing Normal University, College of Life Sciences, Beijing, P. R. China
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  • Feifei Huang,

    1. Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing Normal University, College of Life Sciences, Beijing, P. R. China
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  • Shujun Dong,

    1. Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing Normal University, College of Life Sciences, Beijing, P. R. China
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  • Lin Li,

    1. National Institute of Biological Sciences, Beijing, P. R. China
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  • Ping Gao,

    1. Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing Normal University, College of Life Sciences, Beijing, P. R. China
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  • Heping Zhao,

    1. Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing Normal University, College of Life Sciences, Beijing, P. R. China
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  • Yingdian Wang,

    1. Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing Normal University, College of Life Sciences, Beijing, P. R. China
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  • Shengcheng Han

    Corresponding author
    1. Beijing Key Laboratory of Gene Resource and Molecular Development, Beijing Normal University, College of Life Sciences, Beijing, P. R. China
    • Correspondence: Dr. Shengcheng Han, Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing 100875, P. R. China

      E-mail: schan@bnu.edu.cn

      Fax: +86-10-58807720

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Abstract

Inositol 1,4,5-trisphosohate (IP3) and its receptors play a pivotal role in calcium signal transduction in mammals. However, no homologs of mammalian IP3 receptors have been found in plants. In this study, we isolated the microsomal fractions from rice cells in suspension culture and further obtained putative IP3-binding proteins by heparin-agarose affinity purification. The IP3-binding activities of these protein fractions were determined by [3H] IP3-binding assay. SDS-PAGE and MS analysis were then performed to characterize these proteins. We have identified 297 proteins from the eluates of heparin-agarose column chromatography, which will provide insight into the IP3 signaling pathways in plants. All MS data have been deposited in the ProteomeXchange with identifier PXD000763 (http://proteomecentral.proteomexchange.org/dataset/PXD000763).

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