Systematic analysis of a simple adaptor protein PDZK1: ligand identification, interaction and functional prediction of complex.
PDZK1 is a simple adaptor protein with four protein interaction PDZ domains, but without any other known functional domains. Here, we used yeast two-hybrid screening of a random peptide library and high-throughput validation screening of a specialized PDZ ligand candidate library to systematically and comprehensively identify PDZK1 ligands. The potential functional associations of the ligands were predicted by functional annotations from a MILANO literature search and subcellular localizations. The ligands were considered more likely to be functionally associated if they had similar patterns of functions or closely related functions. For some functionally associated ligand pairs, interaction with one ligand was found to be influenced by another ligand in a yeast three-hybrid system. Many G-protein signaling pathway-related proteins were found to interact with PDZK1, and they were likely to be functionally associated with transporters based on their closely related functions. This strategy can be extended to the study of other adaptor proteins that contain peptide-binding domains.
Hu
Siqi
S
National Key Laboratory of Medical Molecular Biology, Department of Physiology and Pathophysiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences/School of Basic Medicine, Peking Union Medical College, Beijing, China.
Song
Eli
E
Tian
Rui
R
Ma
Sucan
S
Yang
Tao
T
Mu
Yi
Y
Li
Yuan
Y
Shao
Chen
C
Gao
Shijuan
S
Gao
Youhe
Y
D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D015723:Gene Library; D008024:Ligands; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D054731:PDZ Domains; D019151:Peptide Library; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary; D020798:Two-Hybrid System Techniques
D008024:Ligands; D008565:Membrane Proteins; C509767:Pdzk1ip1 protein, mouse; D019151:Peptide Library