Molecular cloning of the cDNA encoding pp36, a tyrosine-phosphorylated adaptor protein selectively expressed by T cells and natural killer cells.
Activation of T and natural killer (NK) cells leads to the tyrosine phosphorylation of pp36 and to its association with several signaling molecules, including phospholipase Cgamma-1 and Grb2. Microsequencing of peptides derived from purified rat pp36 protein led to the cloning, in rat and man, of cDNA encoding a T- and NK cell-specific protein with several putative Src homology 2 domain-binding motifs. A rabbit antiserum directed against a peptide sequence from the cloned rat molecule recognized tyrosine phosphorylated pp36 from pervanadate-treated rat thymocytes. When expressed in 293T human fibroblast cells and tyrosine-phosphorylated, pp36 associated with phospholipase Cgamma-1 and Grb2. Studies with GST-Grb2 fusion proteins demonstrated that the association was specific for the Src homology 2 domain of Grb-2. Molecular cloning of the gene encoding pp36 should facilitate studies examining the role of this adaptor protein in proximal signaling events during T and NK cell activation.
Weber
J R
JR
Department of Medicine and Rosalind Russell Arthritis Center, University of California, San Francisco, California 94143, USA.
Orstavik
S
S
Torgersen
K M
KM
Danbolt
N C
NC
Berg
S F
SF
Ryan
J C
JC
Taskén
K
K
Imboden
J B
JB
Vaage
J T
JT
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003857:Deoxyuridine / Q000031:analogs & derivatives* / Q000737:chemistry; D051380:GRB2 Adaptor Protein; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D007694:Killer Cells, Natural / Q000276:immunology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D051966:Phospholipase C gamma; D010750:Phosphoproteins / Q000737:chemistry; D011412:Propanolamines / Q000737:chemistry*; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000276:immunology; D017422:Sequence Analysis, DNA; D013601:T-Lymphocytes / Q000276:immunology*; D013950:Thymus Gland / Q000502:physiology; D010738:Type C Phospholipases / Q000378:metabolism; D018909:src Homology Domains / Q000235:genetics
D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; C495950:Grb2 protein, rat; D007527:Isoenzymes; D010446:Peptide Fragments; D010750:Phosphoproteins; D011412:Propanolamines; D011506:Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; C081648:5'-((N-(2-decanoylamino-3-hydroxy-3-phenylpropyloxycarbonyl)glycyl)amino)-5'-deoxyuridine; D010738:Type C Phospholipases; D051966:Phospholipase C gamma; D003857:Deoxyuridine