Tyrosine phosphorylation of a 66,000 M/sub r/ soluble protein in lectin-activated human peripheral blood T lymphocytes
Protein phosphorylation was studied in human T lymphocytes stimulated with the mitogenic lectins phytohemagglutinin (PHA) and concanavalin A (Con A). The T lymphocytes were prepared from the venous blood of normal volunteers, their intracellular ATP pools were labeled with (/sup 32/P)orthophosphate, and protein phosphorylation was assayed in the soluble fraction by two-dimensional gel electrophoresis and autoradiography. When lymphocytes stimulated with PHA or Con A were compared to unstimulated control cells, there was a general increase in protein phosphorylation and the specific phosphorylation of a soluble protein with M/sub r/ = 64.9 to 69 KD and pI = 5.6 to 5.8. Phosphorylation of this protein, designated TPP-66, was observed as early as 2 min after the addition of lectin with a gradual increase in the level of phosphorylation over the next 120 min. In the majority of experiments, there was no phosphorylation seen in the unstimulated lymphocytes: however, in some experiments, there was appreciable phosphorylation, which was seen beginning 60 min after the labeling period. When the TPP-66 spot from stimulated lymphocytes was excised from gels, was eluted, and was subjected to limited base hydrolysis followed by single-dimension high voltage electrophoresis, the major phosphorylated residue migrated with phosphotyrosine. In some experiments, there was phosphorylation of serine residues in both the stimulated and control cells; tyrosine phosphorylation was never seen in the unstimulated cell population. These data suggest that, like other stimulae for cell growth, the induction of lymphocyte growth by lectins is associated with the activation of a tyrosine-specific kinase.
- Research Organization:
- Washington Univ. School of Medicine, St. Louis, MO
- OSTI ID:
- 5448317
- Journal Information:
- J. Immunol.; (United States), Vol. 136:11
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
LYMPHOCYTES
STIMULATION
PHOSPHOTRANSFERASES
ENZYME INDUCTION
TYROSINE
PHOSPHORYLATION
ATP
AUTORADIOGRAPHY
BLOOD
CONCANAVALIN
ELECTROPHORESIS
LABELLING
MAN
PHOSPHORUS 32
PHYTOHEMAGGLUTININ
TRACER TECHNIQUES
AGGLUTININS
AMINO ACIDS
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD CELLS
BODY FLUIDS
CARBOHYDRATES
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
ENZYMES
GENE REGULATION
HEMAGGLUTININS
HYDROXY ACIDS
ISOTOPE APPLICATIONS
ISOTOPES
LECTINS
LEUKOCYTES
LIGHT NUCLEI
MAMMALS
MATERIALS
MITOGENS
MUCOPROTEINS
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYSACCHARIDES
PRIMATES
PROTEINS
RADIOISOTOPES
SACCHARIDES
SOMATIC CELLS
TRANSFERASES
VERTEBRATES
551001* - Physiological Systems- Tracer Techniques