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Studies on interaction of insulin and insulin receptor in rat liver cell membranes

Journal Article:

Abstract

Rat liver was homogenized with a Polytron PT 20 ST and fractionated by differential centrifugation. Prepared plasma membranes (100 ..mu..g protein) were incubated with enzymatically iodinated /sup 125/I-insulin (0.3 ng, specific activity 107 ..mu..Ci/..mu..g) in 25 mM Tris-HCl buffer, pH 7.5, containing 0.9% NaCl and 1% bovine serum albumin. The 12,000xg- and 17,000xg-sediments obtained after subfractionation of liver homogenates showed almost equally high specific binding activity with /sup 125/I-insulin and less activity was detected in the 600 g-, 5,000 g- and 40,000 g- sediments and the 40,000 g- supernatant. Specific binding of insulin with the membrane fraction was time-, temperature- and ionic strength-dependent. The highest binding was obtained under conditions in which the membrane fraction was incubated with insulin for 24 hours at 4/sup 0/C in the buffer containing 1 M NaCl. Under these conditions, specific binding of /sup 125/I-insulin was 26.8% of the total radioactivity. The effect of native insulin on the binding of /sup 125/I-insulin with the membrane fraction was studied in the range of 0--6.4 x 10/sup 5/ ..mu..U/ml of unlabeled insulin and a distinct competitive displacement of /sup 125/I-insulin with native insulin was observed between 10 and 10/sup 4/ ..mu..U/ml. Kinetic studies by Scatchard plot analysis  More>>
Authors:
Sakai, Y; Hara, H; Kawate, R; Kawasaki, T [1] 
  1. Hiroshima Univ. (Japan). School of Medicine
Publication Date:
Jul 01, 1975
Product Type:
Journal Article
Reference Number:
AIX-07-273201; EDB-77-027159
Resource Relation:
Journal Name: Nippon Naibumpi Gakkai Zasshi; (Japan); Journal Volume: 51:7
Subject:
59 BASIC BIOLOGICAL SCIENCES; CELL MEMBRANES; INSULIN; METABOLISM; AFFINITY; IODINE 125; LIVER; RADIOPHARMACEUTICALS; RATS; TRACER TECHNIQUES; ANIMALS; BETA DECAY RADIOISOTOPES; BODY; CELL CONSTITUENTS; DAYS LIVING RADIOISOTOPES; DIGESTIVE SYSTEM; DRUGS; ELECTRON CAPTURE RADIOISOTOPES; GLANDS; HORMONES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; LABELLED COMPOUNDS; MAMMALS; MEMBRANES; NUCLEI; ODD-EVEN NUCLEI; ORGANS; PEPTIDE HORMONES; RADIOACTIVE MATERIALS; RADIOISOTOPES; RODENTS; VERTEBRATES; 550501* - Metabolism- Tracer Techniques
OSTI ID:
7240314
Country of Origin:
Japan
Language:
Japanese
Other Identifying Numbers:
Journal ID: CODEN: NNGZA
Submitting Site:
INIS
Size:
Pages: 601-608
Announcement Date:

Journal Article:

Citation Formats

Sakai, Y, Hara, H, Kawate, R, and Kawasaki, T. Studies on interaction of insulin and insulin receptor in rat liver cell membranes. Japan: N. p., 1975. Web.
Sakai, Y, Hara, H, Kawate, R, & Kawasaki, T. Studies on interaction of insulin and insulin receptor in rat liver cell membranes. Japan.
Sakai, Y, Hara, H, Kawate, R, and Kawasaki, T. 1975. "Studies on interaction of insulin and insulin receptor in rat liver cell membranes." Japan.
@misc{etde_7240314,
title = {Studies on interaction of insulin and insulin receptor in rat liver cell membranes}
author = {Sakai, Y, Hara, H, Kawate, R, and Kawasaki, T}
abstractNote = {Rat liver was homogenized with a Polytron PT 20 ST and fractionated by differential centrifugation. Prepared plasma membranes (100 ..mu..g protein) were incubated with enzymatically iodinated /sup 125/I-insulin (0.3 ng, specific activity 107 ..mu..Ci/..mu..g) in 25 mM Tris-HCl buffer, pH 7.5, containing 0.9% NaCl and 1% bovine serum albumin. The 12,000xg- and 17,000xg-sediments obtained after subfractionation of liver homogenates showed almost equally high specific binding activity with /sup 125/I-insulin and less activity was detected in the 600 g-, 5,000 g- and 40,000 g- sediments and the 40,000 g- supernatant. Specific binding of insulin with the membrane fraction was time-, temperature- and ionic strength-dependent. The highest binding was obtained under conditions in which the membrane fraction was incubated with insulin for 24 hours at 4/sup 0/C in the buffer containing 1 M NaCl. Under these conditions, specific binding of /sup 125/I-insulin was 26.8% of the total radioactivity. The effect of native insulin on the binding of /sup 125/I-insulin with the membrane fraction was studied in the range of 0--6.4 x 10/sup 5/ ..mu..U/ml of unlabeled insulin and a distinct competitive displacement of /sup 125/I-insulin with native insulin was observed between 10 and 10/sup 4/ ..mu..U/ml. Kinetic studies by Scatchard plot analysis of the above results revealed heterogeneity in insulin receptors or receptor sites, one with a high affinity of 10/sup 9/ M/sup -1/ order and the other with a low affinity of 10/sup 8/ M/sup -1/ order. Both affinities were also affected by temperature and ionic strength.}
journal = {Nippon Naibumpi Gakkai Zasshi; (Japan)}
volume = {51:7}
journal type = {AC}
place = {Japan}
year = {1975}
month = {Jul}
}