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A short incubation time in insulin binding experiments leads to disappearance of negative cooperativity in H35 hepatoma cells.

Abstract：

:The effects of different periods of incubation (8 min vs 20 min) on insulin binding kinetics were examined in a H35 hepatoma cell line. Scatchard plots from cells incubated for 8 min were linear (r = 0.987 +/- 0.006), in contrast to curvilinear Scatchard plots from cells incubated for 20 min. Hill plots showed a slope of 1.006 +/- 0.024 for the 8 min incubation, whereas the slope was 0.827 +/- 0.0026 (p less than 0.0005) for the 20 min incubation. TCA precipitation of the medium showed minimal insulin degradation products at 8 min with a significant increase at 20 min (1.38 +/- 0.11% vs. 3.06 +/- 0.37%, p less than 0.0005). Internalized insulin was also significantly increased at 20 min as compared to 8 min incubation (48.9 +/- 5.6% vs. 32.4 +/- 3.0%, p less than 0.0005) These data indicate that after 8 min of incubation no appreciable cooperativity of insulin binding was present, while negative cooperativity was present after 20 min of incubation. As significantly more insulin degradation has taken place after prolonged incubation these data support the hypothesis that insulin degradation leads to negative cooperativity of insulin receptors.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

de Vries CP,van Haeften TW,van der Veen EA

doi

10.1016/s0006-291x(88)81029-5

subject

Has Abstract

pub_date

1988-12-30 00:00:00

pages

1390-5

issue

3

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(88)81029-5

journal_volume

157

pub_type

杂志文章

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