Purification and partial characterization of proteinase inhibitors of equine seminal plasma.
The aims of the study were: 1/ to isolate and identify equine seminal plasma proteinase inhibitors, 2/ to evaluate their inhibitory potential, and 3/ to test a correlation between protein concentration in seminal plasma supernatant (obtained after precipitation with 36% ammonium sulfate) and stallion sexual maturity. Seminal plasma proteins obtained from six stallions were chromatographed in a Superose 12 (FPLC system) column followed by C(18) HPLC reverse-phase. Inhibition of trypsin amidase activity was evaluated in the collected fractions. Active proteins with a molecular mass of 6.3-7.0 KDa were identified using mass spectrometry. The older stallions showed a reduction in total seminal plasma protein concentration, but had similar concentrations of proteinase inhibitors (0.28+/-0.10 mg/ml) in seminal plasma supernatant. Different proteinase inhibitor isoforms were found in semen of all stallions which suggests that the isoforms may be used as biomarkers of individual animals.
Vasconcelos
André Belico
AB
Institute of Veterinary José Caetano Borges,, University of Uberaba, Uberaba, Minas Gerais, Av. Tutunas 720, Postal code 38061-500, Brazil. andre_belico@yahoo.com.br
Santos
Alexandre Martins Costa
AM
Oliveira
Jamil Silvano
JS
Lagares
Monique de Albuquerque
Mde A
Santoro
Marcelo Matos
MM
D000375:Aging; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006736:Horses / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D011480:Protease Inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D012661:Semen / Q000737:chemistry*; D029589:Seminal Plasma Proteins / Q000032:analysis / Q000302:isolation & purification; D015842:Serine Proteinase Inhibitors / Q000302:isolation & purification
D011480:Protease Inhibitors; D029589:Seminal Plasma Proteins; D015842:Serine Proteinase Inhibitors