Separation and identification of carp pituitary proteins and glycoproteins
H. Ryšlavá, M. Janatová, G. Čalounová, I. Selicharová, J. Barthová, T. Barthhttps://doi.org/10.17221/4232-CJASCitation:Ryšlavá H., Janatová M., Čalounová G., Selicharová I., Barthová J., Barth T. (2005): Separation and identification of carp pituitary proteins and glycoproteins. Czech J. Anim. Sci., 50: 430-437.
Carp pituitary proteins and glycoproteins were separated by the combination of immobilized metal affinity chromatography (IMAC) and affinity chromatography on Con A-Sepharose. The protein fractions were analysed by SDS-PAGE. Luteinizing hormone (in the α1β and α2β forms), growth hormone, free α1 subunit and β subunit of thyroid-stimulating hormone were identified by N-terminal amino acid sequencing. N-linked oligosaccharide chains of thyroid-stimulating hormone (β TSH) were separated after fluorescent labelling on a GlycoSep N column and treated by exoglycosidases. Among the saccharide components, complex and hybrid structures terminating SO4-GalNAc-GlcNAc-Man-, and high mannose structures, with 1 to 8 mannose units attached to the oligosaccharide core (GlcNAc)2(Man)3, were found.Keywords:
carp hormones; glycoproteins; immobilized metal ion affinity chromatography (IMAC); oligosaccharide chains