Modulation of the steroidogenic related activity according to the design of single-chain bovine FSH analogs

Hila Asraf, Abraham Amsterdam, David Ben-Menahem

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

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.

Original languageEnglish
Pages (from-to)171-181
Number of pages11
JournalGeneral and Comparative Endocrinology
Volume216
DOIs
StatePublished - 15 May 2015

Keywords

  • FSH
  • Glycosylation
  • Single-chain analog
  • Steroidogenesis

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