Novel protein interactors of urokinase-type plasminogen activator receptor.
The urokinase-type plasminogen activator receptor (uPAR) has been implicated in tumor growth and metastasis. The crystal structure of uPAR revealed that the external surface is largely free to interact with a number of proteins. Additionally, due to absence of an intracellular cytoplasmic protein domain, many of the biological functions of uPAR necessitate interactions with other proteins. Here, we used yeast two-hybrid screening of breast cancer cDNA library to identify hSpry1 and HAX1 proteins as putative candidate proteins that interact with uPAR bait constructs. Interaction between these two candidates and uPAR was confirmed by GST-pull down, co-immunoprecipitation assays and confocal microscopy. These novel interactions that have been identified may also provide further evidence that uPAR can interact with a number of other proteins which may influence a range of biological functions.
Mekkawy
Ahmed H
AH
Cancer Research Laboratories, Department of Surgery, St. George Hospital, University of New South Wales, Sydney, NSW 2217, Australia.
De Bock
Charles E
CE
Lin
Zhen
Z
Morris
David L
DL
Wang
Yao
Y
Pourgholami
Mohammad H
MH
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D045744:Cell Line, Tumor; D005260:Female; D015723:Gene Library; D006801:Humans; D047468:Immunoprecipitation; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D055293:Receptors, Urokinase Plasminogen Activator / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D048868:Adaptor Proteins, Signal Transducing; C104759:HAX1 protein, human; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D010750:Phosphoproteins; D055293:Receptors, Urokinase Plasminogen Activator; C486055:SPRY1 protein, human