title: Modulation of the steroidogenic related activity according to the design of single-chain bovine FSH analogs.
abstract: Single-chain (SC) gonadotropins have been genetically engineered to increase the repertoire of analogs for potential use in humans and domestic animals. The major aim of the current study was to examine the steroidogenic related activity of SC FSH analogs carrying structural differences. To address this issue, we designed and expressed three SC bovine FSH analogs in CHO cells: (i) FSHβα in which the tethered subunit domains are linked in tandem; (ii) FSHβCTPα that contains the carboxy terminal peptide (CTP) of the human choriogonadotropin (hCG) β subunit as a spacer, and (iii) FSHβboCTPα in which the linker is derived from a CTP-like sequence (boCTP) decoded from the bovine LHβ DNA. The data suggested that the secretion efficiency of these variants from the transfected cells was unaffected by the presence or absence of the CTP linker, N-glycans were attached to the analogs and the hCGβ-CTP domain in the FSHβCTPα variant was O-glycosylated. In a rat immortalized granulosa cell bioassay the potency of the three variants towards progesterone secretion varied. In immature mice, the analogs increased the ovary weight and induced StAR, Cyp11a (P450scc), Cyp17 (P450c17) and Cyp19 (P450aromatase) transcripts. However, the dose dependence and amplitude of these transcript levels differed in response to FSHβα, FSHβboCTPα and FSHβCTPα. Collectively, these data suggest that the design of the FSH analog can modulate the bioactivity in vitro and in vivo. A systematic analysis of receptor activation with ligands carrying structural differences may identify new regulatory factor/s involved in the pleiotropic FSH activity.
authors: Department of Clinical Biochemistry and Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
authors: Hila
authors: H
authors: Asraf
authors: Department of Molecular and Cellular Biology, Weizmann Institute of Science, Rehovot, Israel.
authors: Abraham
authors: A
authors: Amsterdam
authors: Department of Clinical Biochemistry and Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel. Electronic address: dbm@bgu.ac.il.
authors: David
authors: D
authors: Ben-Menahem
mesh_terms: D000818:Animals; D001141:Aromatase / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells; D002417:Cattle; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D015195:Drug Design; D005260:Female; D005640:Follicle Stimulating Hormone / Q000737:chemistry* / Q000494:pharmacology*; D006031:Glycosylation; D006107:Granulosa Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008813:Mice, Inbred ICR; D010053:Ovary / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism*; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: D010750:Phosphoproteins; D012333:RNA, Messenger; D000098682:Steroidogenic Acute Regulatory Protein; D005640:Follicle Stimulating Hormone; D001141:Aromatase; D013254:Steroid 17-alpha-Hydroxylase; D002786:Cholesterol Side-Chain Cleavage Enzyme
keywords: FSH; Glycosylation; Single-chain analog; Steroidogenesis