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Identification of actin as an estradiol 17‐β‐stimulated protein in the human placenta
Author(s) -
Sudha S.,
Kumari Usha,
Rao Veena S.,
Rao A. Jagannadha
Publication year - 1997
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549700204771
Subject(s) - estrogen , blot , placenta , antiserum , aromatase , stimulation , chemistry , biology , actin , biochemistry , microbiology and biotechnology , endocrinology , gene , fetus , antibody , pregnancy , genetics , cancer , breast cancer , immunology
Addition of estradiol 17‐β to first trimester human placental minces resulted in an increased synthesis of a protein of apparent molecular weight 45 kDa. The specific involvement of estrogen in the stimulation of this protein was established by demonstrating a reduction in the level of this protein by the addition of CGS 16949 A, an inhibitor of aromatase, a key enzyme in the biosynthesis of estradiol 17‐β and ICI 182,780, an estrogen receptor antagonist. The protein was purified to homogeneity and N‐terminal sequencing of two of the internal peptides obtained by enzymatic digestion of the protein, as well as the absence of a free N‐terminal indicated that it could be actin. This was confirmed by Western blotting using commercially available actin antiserum. The role of estradiol 17‐β in the stimulation of actin synthesis in human placenta was also established by monitoring the quantitative inhibition of DNase I by actin.