Rapid detection of protein tyrosine kinase activity in recombinant yeast expressing a universal substrate.
Yeast two-hybrid systems are powerful proteomics tools for the discovery of protein-protein interactions. However, these systems are typically unable to detect interactions dependent on post-translational modifications such as tyrosine phosphorylation. We report a novel yeast tribrid system that expresses a potentially universal protein tyrosine kinase (PTK) substrate to detect diverse PTKs. Validation with the oncogenic kinases v-Abl and v-Src, which exhibit divergent substrate specificities, demonstrated significant potential for cloning PTKs en masse from cDNA libraries.
Clark
Daniel D
DD
Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
Peterson
Blake R
BR
D000818:Animals; D017930:Genes, Reporter; D015394:Molecular Structure; D015688:Oncogene Protein pp60(v-src) / Q000378:metabolism; D016374:Oncogene Proteins v-abl / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D016374:Oncogene Proteins v-abl; D011993:Recombinant Fusion Proteins; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src)