Generation of a lentiviral vector system to efficiently express bioactive recombinant human prolactin hormones. (1st January 2020)

• Record Type: Journal Article
• Authors: Alarcón, Hernán; Bonzon-Kulichenko, Elena; Peinado, Rocío; Lim, Filip; Vázquez, Jesús; Rodríguez, Antonio
• Is Part Of: Molecular and cellular endocrinology. Volume 499(2019)
• Publication Date: 2020-01-01
• Subjects: Prolactin -- Lentiviral vectors -- Recombinant protein expression -- 2A peptide; PRL prolactin -- PRL-R prolactin receptor -- hPRL human prolactin -- hPRL-A mutant human prolactin partial agonist -- hPRL-CA mutant human prolactin competitive antagonist -- GFP green fluorescent protein -- LV lentiviral vector -- P2A 2A peptide; Endocrinology -- Periodicals; Molecular biology -- Periodicals; Cytology -- Periodicals; Endocrinology -- Periodicals; Hormones -- Periodicals; Endocrinologie -- Périodiques; Cytology; Endocrinology; Molecular biology; Periodicals; 573.4
• DOI: 10.1016/j.mce.2019.110605
• Languages: English
• ISSNs: 0303-7207

Abstract

Abstract: The contribution of the pleiotropic hormone Prolactin (PRL) to several physiological and pathological processes is still unknown. To clarify the role of PRL in these processes during the last decade, different human PRL antagonists have been produced to either partially or fully block the wild type hormone activity. In this work, we have cloned these wild type and antagonist sequences in lentivectors (LV) to express them as recombinant self-processing polypeptides by employing a P2A sequence (hPRL-P2A-GFP). We show that these LVs can efficiently transduce and express the hPRL proteins in different cell types and that the P2A sequence does not affect their activities. Additionally, we have tested their activities in paracrine and autocrine cell culture experiments. Our results demonstrate that these recombinant hPRL-P2A proteins are bioactive in both paracrine and autocrine modes, highlighting the potential usefulness of these hPRL-containing LVs for determining the contribution of hPRL to different biological processes. Graphical abstract: Image 1 Highlights: The PRL-P2A-GFP transgenes are efficiently expressed in mammalian cells. The P2A tag renders the recombinant PRL proteins traceable. Recombinant hormone-producing cells can be identified by GFP expression. The recombinant PRL-P2A hormones exhibit paracrine and autocrine bioactivity.