Screen and identification of proteins interacting with ADAM19 cytoplasmic tail.
ADAM family plays important roles in neurogenesis. The cytoplasmic tail of ADAM19 (ADAM19-CT) contains 193 residues. The presence of two putative SH3 ligand-binding sites suggests potential interactions with cytosolic proteins, which could be possibly linked to the functions of ADAM19. To address these issues, a yeast two-hybrid screen was performed in human fetal brain cDNA library to isolate proteins that interact with the cytoplasmic tail of ADAM19. Four proteins were obtained, ArgBP1, beta-cop, ubiquitin and a novel protein. GST-Pulldown assay has confirmed the interaction between AdAM19 and ArgBP1. By constructing series of deletion mutants of ADAM19-CT and ArgBP1 respectively, the interaction regions have been identified. They are the SH3 binding sites in ADAM19-CT and the P4 region in ArgBP1. And the interaction is specific. ArgBP1 does not bind to ADAM22, ADAM29 or ADAM9 (mouse). ArgBP1 may be the key protein, which accounts for the physiological function of ADAM19.
Huang
Li
L
Institute of Genetics, Fudan University, Shanghai 200433, PR. China.
Feng
Libing
L
Yang
Limin
L
Zhou
Weiguo
W
Zhao
Shouyuan
S
Li
Changben
C
D051722:ADAM Proteins; D048868:Adaptor Proteins, Signal Transducing; D001665:Binding Sites; D001923:Brain Chemistry; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism; D020756:Coatomer Protein / Q000302:isolation & purification / Q000378:metabolism; D019483:Disintegrins; D005333:Fetus; D015723:Gene Library; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D008666:Metalloendopeptidases; D045726:Metalloproteases; D009124:Muscle Proteins / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D025801:Ubiquitin / Q000302:isolation & purification / Q000378:metabolism; D018909:src Homology Domains
C096831:ABI2 protein, human; D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D020756:Coatomer Protein; D019483:Disintegrins; D008565:Membrane Proteins; D009124:Muscle Proteins; D011506:Proteins; D025801:Ubiquitin; D045726:Metalloproteases; D051722:ADAM Proteins; C411902:ADAM19 protein, human; C096116:Adam19 protein, mouse; D008666:Metalloendopeptidases