Turnover of insulin receptors in cells transfected with human insulin receptor cDNA

1989 ◽  
Vol 17 (1) ◽  
pp. 197-198
Author(s):  
ROSALIND H. GANDERTON ◽  
JONATHAN WHITTAKER ◽  
KENNETH SIDDLE
Keyword(s):  
Insulin Receptor ◽  
Human Insulin ◽  
Insulin Receptors ◽  
Human Insulin Receptor ◽  
Receptor Cdna ◽  
In Cells

scholarly journals Anti-(insulin receptor) monoclonal antibody-stimulated tyrosine phosphorylation in cells transfected with human insulin receptor cDNA

1990 ◽  
Vol 268 (3) ◽  
pp. 615-620 ◽  
Author(s):  
N P J Brindle ◽  
J M Tavare ◽  
M Dickens ◽  
J Whittaker ◽  
K Siddle
Keyword(s):  
Insulin Receptor ◽  
Tyrosine Phosphorylation ◽  
Human Insulin ◽  
Maximum Level ◽  
Absolute Level ◽  
Insulin Receptor Tyrosine Kinase ◽  
Insulin Mimetic ◽  
Human Insulin Receptor ◽  
Receptor Cdna ◽  
Receptor Antibodies

The effects of insulin and anti-(insulin receptor) monoclonal antibodies on tyrosine phosphorylation were investigated in fibroblasts transfected with human insulin receptor cDNA (NIH 3T3HIR3.5 cells) using anti-phosphotyrosine immunoblotting. Insulin increased levels of tyrosine phosphorylation in two major proteins of molecular mass 97 kDa (pp97, assumed to be the insulin receptor beta-subunit) and 185 kDa (pp185). Insulin-mimetic anti-receptor antibodies also stimulated tyrosine phosphorylation of both pp97 and pp185. The observation of antibody-stimulated pp97 phosphorylation, as detected by immunoblotting, is in contrast with previous data which failed to show receptor autophosphorylation in NIH 3T3HIR3.5 cells labelled with [32P]P1. The effect of insulin on pp97 was maximal within 1 min, but the response to antibody was apparent only after a lag of 1-2 min and rose steadily over 20 min. The absolute level of antibody-stimulated phosphorylation of both pp97 and pp185 after 20 min was only about 20% of the maximum level induced by equivalent concentrations of insulin, even at concentrations of antibody sufficient for full occupancy of receptors. Another insulin-mimetic agent, wheat-germ agglutinin, stimulated receptor autophosphorylation with kinetics similar to those produced by the antibody. It is suggested that the relatively slow responses to both agents may be a function of the dependence on receptor cross-linking. These data are consistent with a role for the insulin receptor tyrosine kinase activity in the mechanism of action of insulin-mimetic anti-receptor antibodies.

The Human Insulin Receptor Cdna: A New Tool to study the Function of this Receptor

1987 ◽  
Vol 7 (1-4) ◽  
pp. 377-404
Author(s):  
Eric Clauser ◽  
Leland Ellis ◽  
David Morgan ◽  
Marc Edery ◽  
Richard A. Roth ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
Human Insulin ◽  
Human Insulin Receptor ◽  
Receptor Cdna

Human insulin receptor radioimmunoassay: applicability to insulin-resistant states

1989 ◽  
Vol 257 (3) ◽  
pp. E451-E457 ◽  
Author(s):  
V. Pezzino ◽  
V. Papa ◽  
V. Trischitta ◽  
A. Brunetti ◽  
P. A. Goodman ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
Human Insulin ◽  
Insulin Receptors ◽  
Insulin Resistant ◽  
Human Insulin Receptor ◽  
Igf I ◽  
Rabbit Polyclonal Antibody ◽  
Specific Activities ◽  
Triton X ◽  
Major Target

A radioimmunoassay of the human insulin receptor was developed employing a potent rabbit polyclonal antibody to the human insulin receptor and a highly purified human placental insulin receptor preparation. The receptor, obtained by sequential affinity chromatography with insulin receptor monoclonal antibody-agarose and wheat germ agglutinin-agarose, was radiolabeled with 125I-Bolton-Hunter reagent at specific activities of 2,100-3,300 Ci/mmol. Over 75% of this ligand was immunoprecipitable with the polyclonal antireceptor antibody and remained immunoprecipitable for greater than 45 days. The assay was sensitive to unlabeled receptor concentrations as low as 0.2 ng/0.5 ml; unlabeled insulin did not cross-react and unlabeled insulin-like growth factor (IGF)-I receptor cross-reacted weakly. The radioimmunoassay was applicable to the measurement of insulin receptors in tissues and cells that were extracted by solubilization in 1% Triton X-100; no purification of the extracted receptor was necessary. Of the three major target tissues for insulin action studied, liver had the highest concentration of receptors (47.6 ng/mg protein); fat and muscle had lower levels. Other studies with the radioimmunoassay indicated that insulin receptors were decreased both in monocytes from obese hyperinsulinemic subjects and in fibroblasts from patients with leprechaunism.

The human insulin receptor cDNA: The structural basis for hormone-activated transmembrane signalling

Cell ◽  
1985 ◽  
Vol 40 (4) ◽  
pp. 747-758 ◽  
Author(s):  
Yousuke Ebina ◽  
Leland Ellis ◽  
Kurt Jarnagin ◽  
Marc Edery ◽  
Laszlo Graf ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
Human Insulin ◽  
Structural Basis ◽  
Transmembrane Signalling ◽  
Human Insulin Receptor ◽  
Receptor Cdna

scholarly journals High-level expression of human insulin receptor cDNA in mouse NIH 3T3 cells.

1987 ◽  
Vol 84 (15) ◽  
pp. 5237-5241 ◽  
Author(s):  
J. Whittaker ◽  
A. K. Okamoto ◽  
R. Thys ◽  
G. I. Bell ◽  
D. F. Steiner ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
Human Insulin ◽  
3T3 Cells ◽  
High Level Expression ◽  
Human Insulin Receptor ◽  
Receptor Cdna ◽  
High Level ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

scholarly journals Dynamic association of human insulin receptor with lipid rafts in cells lacking caveolae

EMBO Reports ◽  
2002 ◽  
Vol 3 (1) ◽  
pp. 95-100 ◽  
Author(s):  
Saara Vainio ◽  
Sanna Heino ◽  
Jan‐Eric Månsson ◽  
Pam Fredman ◽  
Esa Kuismanen ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
Lipid Rafts ◽  
Human Insulin ◽  
Human Insulin Receptor ◽  
Dynamic Association ◽  
In Cells
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