Synthesis of specific estradiol-induced protein by surviving rat uteri and cell-free uterine extracts

Dalia Sömjen, Alvin M. Kaye, Hans R. Lindner

Research output: Contribution to journalArticlepeer-review

Abstract

A cell-free system was developed for the synthesis of the specific 'induced protein' (IP) which is stimulated in the rat uterus within the first hour after treatment with estradiol-17β. Surviving uteri or uterine extracts were incubated with [3H]-labeled (estrogen-treated) or with [14C]- or [3S]-labeled (controls) leucine, glutamic acid or methionine, and the 150,000 gav supernatant solution was analyzed by electrophoresis on cellulose acetate. IP was detected by determining the isotope incorporation ratio. The 12,000 gmax post-mitochondrial supernatant suspension from uteri of 10- or 20-day old rats, prepared 1 h after injection of estradiol-17β (4 or 5 μg/rat), was found to synthesize IP. Treatment of surviving uteri with 30 nM estradiol-17β for l h at 37°C also permitted the extraction of a post-mitochondrial supernatant suspension capable of synthesizing IP. The electrophoretic distribution of radiolabeled soluble proteins was different in the cell-free system and surviving uteri. However, both preparations showed a single protein band with increased incorporation of tritiated amino acid after estradiol treatment. This band had an electrophoretic mobility relative to bovine serum albumin of approximately 1.1, characteristic of IP. Appearance of this band was prevented by addition of antibodies to estradiol or of cordycepin during incubation of the uteri with estradiol. The induction of IP in surviving uteri followed by IP synthesis in a cell-free system provides a versatile model for the more detailed analysis of the regulation of specific protein synthesis by estradiol.

Original languageEnglish
Pages (from-to)77-88
Number of pages12
JournalMolecular and Cellular Endocrinology
Volume1
Issue number2
DOIs
StatePublished - Apr 1974

Keywords

  • actions uterus
  • induced protein
  • oestradiol-17β
  • protein synthesis

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