Journal article
Isolation and characterization of FSH and LH from pituitary glands of Atlantic halibut (Hippoglossus hippoglossus L.)
General and comparative endocrinology, Vol.131(2), pp.97-105
04/2003
Handle:
https://hdl.handle.net/2376/106226
PMID: 12679086
Abstract
Two gonadotropins (GtH), follicle-stimulating hormone (FSH) and luteinizing hormone (LH), were isolated and characterized from pituitary glands of Atlantic halibut (Hippoglossus hippoglossus L.). Glycoproteins were extracted in 40% ethanol followed by precipitation in 85% ethanol. Subsequently, glycoproteins were fractionated by ion-exchange chromatography on a Whatman DE-52 column using a stepwise gradient of ammonium bicarbonate (50-1000 mM). Intact FSH and LH were finally purified on rpHPLC using an AsahiPak C4P-50 column with an acetonitrile gradient (10-60%). SDS-PAGE showed a molecular mass of 33 and 32 kDa for intact FSH and LH, respectively. Final purification of subunits was performed by a subsequent purification step on rpHPLC using a Phenomenex Jupiter C18 column with an acetonitrile gradient (10-60%). FSHbeta, LHbeta, and the common alpha subunit showed molecular masses of 25, 24, and 19 kDa, respectively. Subunit identity was confirmed by N-terminal amino acid sequencing. Intact FSH and LH showed gonadotropic activity by stimulating release of 11-ketotestosterone from turbot (Scophthalmus maximus L.) testicular tissue in vitro. This provides the first purification of two distinct GtHs from an evolutionary advanced pleuronectiform teleost.
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Details
- Title
- Isolation and characterization of FSH and LH from pituitary glands of Atlantic halibut (Hippoglossus hippoglossus L.)
- Creators
- Finn-Arne Weltzien - Institute of Marine Research, Austevoll Aquaculture Research Station, 5392 Storebø, Norway. f.a.weltzien@bio.uio.noBirgitta NorbergPenny Swanson
- Publication Details
- General and comparative endocrinology, Vol.131(2), pp.97-105
- Academic Unit
- UNKNOWN
- Publisher
- United States
- Identifiers
- 99900546951001842
- Language
- English
- Resource Type
- Journal article