Direct protein-protein interaction between PLCgamma1 and the bradykinin B2 receptor--importance of growth conditions.
Recently, we have described a novel protein-protein interaction between the G-protein coupled bradykinin B2 receptor and tyrosine phosphatase SHP-2 via an immunoreceptor tyrosine-based inhibition motif (ITIM) sequence located in the C-terminal part of the B2 receptor and the Src homology (SH2) domains of SHP-2. Here we show that phospholipase C (PLC)gamma1, another SH2 domain containing protein, can also interact with this ITIM sequence. Using surface plasmon resonance analysis, we observed that PLCgamma1 interacted with a peptide containing the phosphorylated form of the bradykinin B2 receptor ITIM sequence. In CHO cells expressing the wild-type B2 receptor, bradykinin-induced transient recruitment and activation of PLCgamma1. Interestingly, this interaction was only observed in quiescent and not in proliferating cells. Mutation of the key ITIM residue abolished this interaction with and activation of PLCgamma1. Finally we also identified bradykinin-induced PLCgamma1 recruitment and activation in primary culture renal mesangial cells.
Duchene
Johan
J
Inserm U388, IFR 31, Hopital Rangueil, TSA 50032, 31059 Toulouse Cedex 9, France.
Chauhan
Sharmila D
SD
Lopez
Frédéric
F
Pecher
Christiane
C
Estève
Jean-Pierre
JP
Girolami
Jean-Pierre
JP
Bascands
Jean-Loup
JL
Schanstra
Joost P
JP
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D008969:Molecular Sequence Data; D051966:Phospholipase C gamma; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D043782:Receptor, Bradykinin B2 / Q000737:chemistry* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance / Q000379:methods; D010738:Type C Phospholipases / Q000737:chemistry* / Q000378:metabolism*
D043782:Receptor, Bradykinin B2; D010738:Type C Phospholipases; D051966:Phospholipase C gamma