Proteome identification of binding-partners interacting with cell polarity protein Par3 in Jurkat cells.
The evolutionarily conserved cell polarity protein Par3, a scaffold-like PDZ-containing 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 combining the 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-alpha4 and proteasome activators PA28beta and PA28gamma--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.
Zhou
Ying
Y
School of Life Science, East China Normal University, Shanghai 200062, China.
Fang
Longhou
L
Du
Dan
D
Zhou
Wenchao
W
Feng
Xiujing
X
Chen
Jiwu
J
Zhang
Zhe
Z
Chen
Zhengjun
Z
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001324:Autoantigens / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D016764:Cell Polarity / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D004285:Dogs; D006801:Humans; D019169:Jurkat Cells; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D054731:PDZ Domains; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D020543:Proteome / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D028901:alpha Karyopherins / Q000235:genetics / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D001324:Autoantigens; D018797:Cell Cycle Proteins; D017931:DNA Primers; C106306:KPNA3 protein, human; C099558:Ki antigen; D008565:Membrane Proteins; D009124:Muscle Proteins; C509689:PARD3 protein, human; D020543:Proteome; D011993:Recombinant Fusion Proteins; D028901:alpha Karyopherins; C486207:PSME2 protein, human; D046988:Proteasome Endopeptidase Complex