Faculty Collaboration Database

Characterization of an interleukin-2 (IL-2)-induced tyrosine phosphorylated 116-kDa protein associated with the IL-2 receptor beta-subunit. J Biol Chem 1993 Oct 25;268(30):22765-70

Date

10/25/1993

Pubmed ID

7693677

Scopus ID

2-s2.0-0027440218 (requires institutional sign-in at Scopus site)   23 Citations

Abstract

In this paper we have extended previous results on interleukin-2 receptor (IL2-R) signal transduction and focused on the interleukin-2 (IL-2)-stimulated tyrosine phosphorylation of a 116-kDa protein (p116) observed in IL-2 responsive cells. This protein exhibited rapid and transient phosphorylation kinetics in both human T-lymphocytes and the YT cell line, attaining maximum tyrosine phosphorylation within 5 min of stimulation with IL-2. Tyrosine phosphorylated p116 co-purified with activated IL-2 receptor beta-chain (IL2-R beta) when IL2-R complexes were covalently stabilized with the membrane-permeable cleavable cross-linking agent dithiobis(succimidyl propionate) prior to detergent cell lysis and immunoprecipitation with monoclonal anti-IL2-R beta antibodies. Under these conditions comparable amounts of tyrosine-phosphorylated p116 were immunoprecipitated with either anti-IL2-R beta antibodies or anti-phosphotyrosine antibodies, suggesting that a major portion of tyrosine phosphorylated p116 is associated with the IL2-R beta subunit. Furthermore, unphosphorylated p116 was also associated with unactivated IL2-R beta, based on the observation that p116 from unstimulated YT cells underwent tyrosine phosphorylation in IL2-R beta immune-complex tyrosine kinase assay as demonstrated by anti-phosphotyrosine immunoblotting. The presence of tyrosine kinase activity in affinity-purified IL2-R beta complexes supports the notion of a preformed receptor-kinase complex. The co-association of both p116 and tyrosine kinase activity with the IL2-R beta supports the critical role of the beta-chain in IL2-R signal transduction and suggests that p116 may have a role in the dynamics of IL2-R activation.

Author List

Kirken RA, Rui H, Evans GA, Farrar WL

MESH terms used to index this publication - Major topics in bold

Cell Line
Cells, Cultured
Electrophoresis, Polyacrylamide Gel
Humans
Interleukin-2
Kinetics
Lymphocyte Activation
Macromolecular Substances
Molecular Weight
Phosphoproteins
Phosphorylation
Phosphotyrosine
Receptor Protein-Tyrosine Kinases
Receptors, Interleukin-2
T-Lymphocytes
Tyrosine