1. Explorations of the Insulinlike and Growth-Promoting Properties of Somatomedin by Membrane Receptor Assays
- Author
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Louis E. Underwood, J J Van Wyk, Raymond L. Hintz, Joel B. Baseman, Robert N. Marshall, and David R. Clemmons
- Subjects
medicine.medical_specialty ,animal structures ,biology ,Chemistry ,Isoelectric focusing ,Somatomedin ,Insulin receptor ,Membrane ,Endocrinology ,Biochemistry ,Sephadex ,Cell surface receptor ,Internal medicine ,medicine ,biology.protein ,Receptor ,Polyacrylamide gel electrophoresis ,hormones, hormone substitutes, and hormone antagonists - Abstract
Publisher Summary This chapter presents the explorations of the insulinlike and growth-promoting properties of somatomedin by membrane receptor assays. A competitive radioreceptor assay using 125 I-labeled insulin and a human placental cell membrane preparation was developed to measure the total insulinlike activity (ILA) during the fractionation of outdated human plasma. After isoelectric focusing, a major peak of ILA focused in a neutral zone and a minor peak in a more basic zone; conversely, more of the bioassayable somatomedin activity focused in the basic zone than in the neutral zone. The basic fraction was further purified by preparative electrophoresis at pH 2.3 on a 15% acrylamide gel containing 4 M urea. After additional chromatographic steps on Sephadex, an arginine-rich peptide was obtained and designated somatomedin C. Somatomedin C labeled with radioactive iodine was found to possess its own primary binding site in human placental cell membranes, chick embryo chondrocyte membranes, and membranes from rat liver, thymus, kidney, muscle, and brain. The somatomedin C receptor was 100 times more sensitive to competitive binding by unlabeled somatomedin than was the insulin receptor and 1000 times less sensitive to competitive inhibition by insulin.
- Published
- 1975
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