Proteome identification of binding-partners interacting with cell polarity protein Par3 in Jurkat cells
Ying Zhou
School of Life Science, East China Normal University, Shanghai 200062, China
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
These authors contributed equally to this work
Search for more papers by this authorLonghou Fang
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
These authors contributed equally to this work
Current address: Department of Medicine, University of California, San Diego, CA 92093, USA
Search for more papers by this authorDan Du
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China
Search for more papers by this authorWenchao Zhou
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China
Search for more papers by this authorXiujing Feng
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China
Search for more papers by this authorJiwu Chen
School of Life Science, East China Normal University, Shanghai 200062, China
Search for more papers by this authorZhe Zhang
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
Search for more papers by this authorCorresponding Author
Zhengjun Chen
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
*Corresponding author: Tel, 86-21-54921081; Fax, 86-21-54921081; E-mail, [email protected]Search for more papers by this authorYing Zhou
School of Life Science, East China Normal University, Shanghai 200062, China
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
These authors contributed equally to this work
Search for more papers by this authorLonghou Fang
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
These authors contributed equally to this work
Current address: Department of Medicine, University of California, San Diego, CA 92093, USA
Search for more papers by this authorDan Du
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China
Search for more papers by this authorWenchao Zhou
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China
Search for more papers by this authorXiujing Feng
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China
Search for more papers by this authorJiwu Chen
School of Life Science, East China Normal University, Shanghai 200062, China
Search for more papers by this authorZhe Zhang
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
Search for more papers by this authorCorresponding Author
Zhengjun Chen
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China
*Corresponding author: Tel, 86-21-54921081; Fax, 86-21-54921081; E-mail, [email protected]Search for more papers by this authorThis work was supported by grants from the National Natural Science Foundation of China (Nos. 30730055, 30700129 and 30623002) and the National Basic Research Program of China (No. 2007CB914504)
Current address: Department of Medicine, University of California, San Diego, CA 92093, USA
Abstract
The evolutionarily conserved cell polarity protein Par3, a scaffold-like PDZreontaining protein, plays a critical role in the establishment and maintenance of epithelial cell polarity. Although the role of Par3 in establishing cell polarity in epithelial cells has been intensively explored, the function of Par3 in hematopoietic cells remains elusive. To address this issue, we generated GST-fusion proteins of Par3 PDZ domains. By combiningthe GST-pull-down approach with liquid chromatography-tandem mass spectrometry, we identified 10 potential novel binding proteins of PDZ domains of Par3 in Jurkat cells (a T-cell line). The interaction of Par3 with three proteins—nuclear transport protein importin-α4 and proteasome activators PA28β and PA28γ—was confirmed using in vitro binding assay, co-immunoprecipitation assay and immunofluorescence microscopy. Our results have the potential to uncover novel functions of the cell polarity protein Par3 in blood cells.
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