Analysis of growth hormone and lactogenic binding sites cross-linked to iodinated human growth hormone

Journal Article · Wed Jun 01 00:00:00 EDT 1983 · Endocrinology; (United States)

DOI:https://doi.org/10.1210/endo-112-6-1980· OSTI ID:5429235

Hughes, J P; Simpson, J S; Friesen, H G

GH (GHR) and lactogenic receptors were analyzed after use of the cross-linking reagent ethylene glycol bis-(succinimidyl succinate) to attach covalently iodinated human GH (hGH) to binding proteins 1) on intact IM-9 lymphocytes, 2) in a partially purified GHR preparation from rabbit liver, and 3) in crude microsomal fractions from rabbit liver, rabbit mammary gland, and rat liver. The latter two microsomal preparations contain primarily lactogenic receptors, whereas in IM-9 lymphocytes and the rabbit liver preparations, GHR predominate. Cross-linked (125I)hGH-receptor complexes were solubilized, reduced, and separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Analysis of proteins cross-linked to (125I)hGH in the microsomal fraction from rabbit liver showed a specifically labeled complex with an estimated molecular weight (mol wt) of 75K. A slightly lower mol wt (71K) was determined for the complex labeled in the purified GHR preparation. In contrast to the relatively low mol wt complexes in rabbit liver, a complex that migrated with an apparent mol wt of 130K was identified in IM-9 lymphocytes. Labeled complexes were identified at 66K from rat liver and 61K from rabbit mammary gland. If it is assumed that hGH contributes 21K to the mol wt of the radiolabeled complexes, then the approximate mol wts of hGH-binding sites are 50-54K from rabbit liver, 109K from IM-9 lymphocytes, 45K from rat liver, and 40K from rabbit mammary gland.

• Research Organization: Department of Physiology, University of Manitoba, Winnipeg, Canada
• OSTI ID: 5429235
• Journal Information: Endocrinology; (United States), Vol. 112:6
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
RECEPTORS
AUTORADIOGRAPHY
ANIMAL GROWTH
HORMONES
IODINE 125
LABELLING
LIVER
LYMPHOCYTES
MAMMARY GLANDS
MAN
MICROSOMES
PREGNANCY
PROTEINS
RABBITS
RATS
ANIMAL CELLS
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DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
GLANDS
GROWTH
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPES
LEUKOCYTES
MAMMALS
MATERIALS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANOIDS
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551001* - Physiological Systems- Tracer Techniques