Collect. Czech. Chem. Commun. 1999, 64, 1510-1516
https://doi.org/10.1135/cccc19991510

Chicken Pituitary Glycoproteins: New Isolation Method

Helena Ryšlaváa, Jana Krešlováa, Jana Barthováa,* and Tomislav Barthb

a Department of Biochemistry, Charles University, Albertov 2030, 128 40 Prague 2, Czech Republic
b Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Prague 6, Czech Republic

Abstract

A new method for isolation of glycoproteins from chicken pituitaries was applied. The procedure consist of chromatography on ConA-Sepharose and by HPLC on S Hyper D and Vydac C4 columns. The hormonal activity of the glycoproteins was tested by determining their stimulatory effect on cAMP or testosterone production. Molecular weights of the products of tryptic cleavage of the hormone were determined using mass spectrometry (MALDI TOF). A comparison of the values obtained with theory shows that the protein is the β-unit of chicken luteinizing hormone.

Keywords: Chicken pituitary; Glycoproteins; Hormones; Gonadotropins; Proteins; Mass spectrometry; Ion-exchange chromatography; HPLC.