Molecular cloning of p125Nap1, a protein that associates with an SH3 domain of Nck.
Binding proteins to the Src homology 3 (SH3) domains of Nck were screened by the use of glutathione S-transferase fusion proteins. Two proteins of 140 and 125 kDa were detected, both of which associated preferentially with the first SH3 domain of Nck. The 125-kDa protein, designated as Nap1 for Nck-associated protein 1, was purified and the corresponding rat cDNA was isolated. The predicted amino acid sequence revealed that p125Nap1 does not contain any known functional motif but shows sequence homology to Hem family gene. Using specific antibodies, p125Nap1 was shown to associate with Nck both in vitro and in intact cells. Further characterization of p125Nap1 may clarify the protein-protein interaction in the downstream signaling of Nck.
Kitamura
T
T
Second Department of Internal Medicine, Kobe University School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Japan.
Kitamura
Y
Y
Yonezawa
K
K
Totty
N F
NF
Gout
I
I
Hara
K
K
Waterfield
M D
MD
Sakaue
M
M
Ogawa
W
W
Kasuga
M
M
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015513:Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D051381:Rats; D014162:Transfection; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; C484362:NCKAP1 protein, human; C078332:Nck protein; C098500:Nckap1 protein, rat; D015513:Oncogene Proteins; D005982:Glutathione Transferase