Affinity liquid chromatography and capillary electrophoresis of seminal plasma proteins.
Interactions of boar, bull, and human seminal plasma proteins with heparin and phosphorylcholine were studied by affinity LC using heparin immobilized to a Toyopearl support. A step gradient elution from 0.15 to 1.50 M NaCl was employed to elute the seminal plasma proteins. Relative amounts of the heparin-binding fraction of seminal plasma proteins (H+) in seminal plasma of three species were determined. Further on, the fraction of seminal plasma proteins interacting with phosphorylcholine-binding proteins (P+) was evaluated. P+ proteins were not found in human seminal plasma and their highest amount was present in bull seminal plasma. A CE method was developed for separation of seminal plasma proteins. Various capillaries and separation conditions were tested; the best resolution was obtained in a bare-silica capillary, with a micellar system consisting of a 0.02 M borate buffer and 0.05 M SDS pH 10.0. The optimized conditions were applied to the identification of the components in boar plasma.
Varilová
Tereza
T
Department of Analytical Chemistry, Faculty of Science, Charles University, Prague, Czech Republic. tereza.varilova@seznam.cz
Seménková
Hana
H
Horák
Pavel
P
Madera
Milan
M
Pacáková
Vera
V
Tichá
Marie
M
Stulík
Karel
K
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006493:Heparin / Q000737:chemistry; D006801:Humans; D008297:Male; D010767:Phosphorylcholine / Q000737:chemistry; D020033:Protein Isoforms / Q000032:analysis; D029589:Seminal Plasma Proteins / Q000032:analysis*
D020033:Protein Isoforms; D029589:Seminal Plasma Proteins; D010767:Phosphorylcholine; D006493:Heparin