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Recombinant expression and characterization of human activin A
R H Schwall1, K Nikolics, E Szonyi
1Department of Developmental Biology, Genentech, Inc., South San Francisco, California 94080.
Molecular Endocrinology (Baltimore, Md.)
|December 1, 1988
Summary
Recombinant activin A, a protein stimulating FSH secretion, was successfully produced. This confirms activin A
Area of Science:
- Endocrinology
- Reproductive Biology
- Molecular Biology
Background:
- Activin stimulates Follicle-Stimulating Hormone (FSH) secretion from anterior pituitary cells.
- Activin is composed of beta-subunits of inhibin, with species A and AB identified.
- Native activin A has been purified and characterized from ovarian follicular fluid.
Purpose of the Study:
- To biosynthetically produce recombinant human activin A.
- To characterize the biochemical properties and biological activities of recombinant activin A.
- To confirm the structure and function of activin A.
Main Methods:
- Construction of stable cell lines expressing the mRNA for the beta A subunit of human inhibin.
- Biosynthesis and purification of recombinant human activin A.
- Assays for FSH secretion in cultured pituitary cells and hemoglobin accumulation in K562 cells.
Main Results:
- Stable cell lines secreted a 24 kDa beta A dimer, identified as recombinant activin A.
- Recombinant activin A stimulated FSH secretion, with activity sensitive to disulfide bond reduction, slow onset, and blocked by actinomycin D.
- Recombinant activin A also stimulated hemoglobin accumulation in K562 cells.
Conclusions:
- Recombinant activin A possesses the biochemical and biological properties of native activin A.
- The study provides independent confirmation of activin A's structure and function.
- This work validates the use of recombinant technology for studying activin's role in reproductive biology.