Abstract
Activin plays an important role in numerous physiological processes such as cell differentiation and remodeling, regeneration and repair of tissues from various organs, angiogenesis, morphogenesis of glandular organs, pluripotency and differentiation of stem cells, cell adhesion and apoptosis. It participates in reproductive processes like embryogenesis, in the expression of Follicle Stimulating Hormone and Luteinizing Hormone and maturation of ovarian follicles and therefore has application in the area of reproduction of vertebrates. Given the economic importance of activin, we develop an efficient and economical method for the production of recombinant human activin A (rACT), using as expression system the yeast Pichia pastoris. rACT showed biological activity as it induced, on submicromolar dose, the increase of ovarian mass and the ovulation process in a mammal model.
Highlights
Activins are homodimeric proteins which are present in three different isoforms [1]-[3]
Expression of Recombinant Human Activin A The supernatant from fermentation (4 L) was dialyzed and concentrated in the hollow fiber system to 0.5 L, the protein content estimated (0.42 mg/mL) and one sample loaded on a 12.5% SDS-PAGE (Figure 1)
All animals of recombinant human activin A (rACT)-treated group presented increased ovarian mass, including that treated with crude protein at higher doses (5 μg/animal/day) with similar results
Summary
Activins are homodimeric proteins which are present in three different isoforms [1]-[3]. Each isoform is constituted by two beta subunits (14 kDa) linked by disulphide bonds [1] [4]. Activins are synthesized in various tissues and its expression levels are elevated in the hypothalamus, pituitary gland, adrenal and placenta [5] [6]. They participate in numerous physiological processes, such as cellular differentiation and remodeling [7]-[9], neural survival [10], angiogenesis [11], tissue repair [12], cell adhesion and apoptosis [6]. Activin A is distributed in spermatocytes, eggs and in various organs during embryogenesis [17] and is important to support granulosa cells survival and proliferation [18]
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