[Screening and identification of interactive proteins of SH2D4A].
SH2D4A is a member of SH2 signaling protein family, which is involved in the signal transduction mediated by protein tyrosine kinase-related receptor, cell growth, proliferation, differentiation, and thereby affects the development of human disorders. To determine the role of SH2D4A in the cell signal transduction pathway, SH2D4A interactive proteins were screened using yeast two-hybrid system, and yeast mating and GST pull-down assays were carried out to further confirm the interaction. We successfully generated a bait protein expression construct-pGBKT7-SH2D4A, screened the human kidney cDNA library, and obtained 46 positive yeast clones. After isolation of positive colonies, DNA sequencing, and sequence alignment analysis with BLAST software, we obtained 5 potential SH2D4A interactive proteins, AZGP1, DAD1, HSD17B10, KAT5, and PKM2, which were predicted by NetPhos 2.0 Server software and were all shown to be phosphorylated tyrosine (pY)-containing proteins except for HSD17B10. KAT5 and HSD17B10 were selected to perform yeast mating and GST pull-down experiments, indicating their direct binding to SH2D4A.
Li
Wei
W
Department of Medical genetics, China Medical University, Shenyang 110001, China. weili19801130@hotmail.com
Li
Ting-Ting
TT
Liu
Hong
H
Zhao
Yan-Yan
YY
D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000378:metabolism; D054392:Adipokines; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000378:metabolism*; D011485:Protein Binding; D012984:Software; D020798:Two-Hybrid System Techniques; D000090203:Zn-Alpha-2-Glycoprotein
D015094:3-Hydroxyacyl CoA Dehydrogenases; D054392:Adipokines; D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins; D006023:Glycoproteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C488891:AZGP1 protein, human; D000090203:Zn-Alpha-2-Glycoprotein; C083331:DAD1 protein, human; C474507:SH2D4A protein, human; C108634:HSD17B10 protein, human