Identification of the insulin receptor tyrosine residues undergoing insulin-stimulated phosphorylation in intact rat hepatoma cells

Journal Article · Tue Jan 05 00:00:00 EST 1988 · J. Biol. Chem.; (United States)

OSTI ID:6837967

Tornqvist, H E; Gunsalus, J R; Nemenoff, R A; Frackelton, A R; Pierce, M W; Avruch, J

Tyr(P)-containing proteins were purified from extracts of insulin-treated rat hepatoma cells (H4-II-E-C3) by antiphosphotyrosine immunoaffinity chromatography. Two major insulin-stimulated, Tyr(P) proteins were recovered: an M/sub r/ 95,000 protein (identified as the insulin receptor ..beta.. subunit by its immunoprecipitation by a patient-derived anti-insulin receptor serum and several anti-insulin receptor (peptide) anti-sera) and an M/sub r/ 180,000 protein. After purification and tryptic digestion of the M/sub r/ 95,000 protein, tryptic peptides containing Tyr (P) were purified by sequential antiphosphotyrosine immunoaffinity, reversed-phase, anion-exchange chromatography. Approximately 80% of all ..beta.. subunit (/sup 32/P)Tyr(P) resides on two tryptic peptides: 50-60% of (/sup 32/P)Tyr(P) is found on the tryptic peptide Asp-Ile-Try-Glu-Thr-Asp-Try-Try-Arg from the tyrosine kinase domain. A second tryptic peptide is located near the carboxyl terminus; this contains 20-30% of ..beta.. subunit (/sup 32/P)Tyr(P) and is identified primarily in a double phosphorylated form. In a summary, the insulin-stimulated tyrosine phosphorylation of the insulin receptor in intact rat hepatoma cells involves at least 6 of the 13 tyrosine residues located on the ..beta.. subunit intracellular extension. These tyrosines are clustered in several domains in a distribution virtually identical to that previously found for partially purified human insulin receptor autophosphorylated in vitro in the presence of insulin.

• Research Organization: Harvard Medical School, Boston, MA (USA)
• OSTI ID: 6837967
• Journal Information: J. Biol. Chem.; (United States), Vol. 263:1
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
RECEPTORS
MOLECULAR STRUCTURE
PURIFICATION
TYROSINE
PHOSPHORYLATION
BLOOD SERUM
HEPATOMAS
INSULIN
ION EXCHANGE CHROMATOGRAPHY
LABELLED COMPOUNDS
LIVER CELLS
PATIENTS
PEPTIDES
PHOSPHORUS 32
TUMOR CELLS
AMINO ACIDS
ANIMAL CELLS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DISEASES
HORMONES
HYDROXY ACIDS
ISOTOPES
LIGHT NUCLEI
MEMBRANE PROTEINS
NEOPLASMS
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
PEPTIDE HORMONES
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
SEPARATION PROCESSES
SOMATIC CELLS
550201* - Biochemistry- Tracer Techniques