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p56lck stably associates with a 115 kDa substrate
P R Srinivas1, J E Hutchcroft, B D Aube
1Department of Medicinal Chemistry and Pharmacognosy, Purdue University, West Lafayette, IN 47907.
Biochemical and Biophysical Research Communications
|July 15, 1992
Summary
Researchers identified a 115 kDa protein (p115) that binds to p56lck. This complex
Area of Science:
- Cellular Biology
- Immunology
- Protein Biochemistry
Background:
- p56lck is a critical kinase in immune cell signaling.
- Understanding p56lck interactions is key to deciphering immune responses.
Purpose of the Study:
- To identify proteins that associate with p56lck.
- To characterize the properties and regulation of the p56lck/p115 complex.
Main Methods:
- Immunoprecipitation using antibodies against p56lck.
- Analysis of protein complexes in whole cell lysates.
- In vitro kinase assays to assess tyrosine phosphorylation.
- Treatment of cells with 12-O-tetradecanoyl phorbol-13-acetate (TPA).
Main Results:
- A 115 kDa protein (p115) was specifically co-immunoprecipitated with p56lck.
- The p56lck/p115 complex demonstrated stability in nonionic detergents.
- p115 undergoes tyrosine phosphorylation in p56lck immune-complex kinase assays.
- TPA treatment reduced subsequent tyrosine phosphorylation of p115.
Conclusions:
- p115 is a novel binding partner of p56lck.
- The p56lck/p115 interaction is robust and occurs in a detergent-stable complex.
- Tyrosine phosphorylation of p115 is regulated, potentially by signaling pathways involving TPA.