Heparin-binding proteins of human seminal plasma: purification and characterization.
Human seminal plasma (HuSP) contains several proteins that bind heparin and related glycosaminoglycans. Heparin binding proteins (HBPs) from seminal plasma have been shown to participate in modulation of capacitation or acrosome reaction and thus have been correlated with fertility in some species. However, these have not been studied in detail in human. The objective of this study was to purify major HBPs from HuSP in order to characterize these proteins. HBPs were isolated by affinity-chromatography on Heparin-Sepharose column, purified by reverse-phase high-performance liquid chromatography (RP-HPLC) and Size-exclusion chromatography and checked for purity on sodium-dodecyl PAGE (SDS-PAGE). Identification of HBPs was done by matrix-assisted laser desorption-ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS). Here we report the purification and identification of seven HBPs in seminal fluid. The major HBPs are lactoferrin and its fragments, semenogelin I fragments, semenogelin II, prostate specific antigen, homolog of bovine seminal plasma-proteins (BSP), zinc finger protein (Znf 169) and fibronectin fragments. In this study we are reporting for the first time the purification and identification of BSP-homolog and Znf 169 from HuSP and classified them as HBPs. Here we report the purification of seven clinically important proteins from human seminal fluid through heparin affinity chromatography and RP-HPLC, in limited steps with higher yield.
Kumar
Vijay
V
Department of Biophysics, All India Institute of Medical Sciences, New Delhi, India.
Hassan
Md Imtaiyaz
MI
Kashav
Tara
T
Singh
Tej P
TP
Yadav
Savita
S
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D007781:Lactoferrin / Q000737:chemistry / Q000302:isolation & purification*; D008297:Male; D012661:Semen / Q000737:chemistry*; D029607:Seminal Vesicle Secretory Proteins / Q000737:chemistry / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D029607:Seminal Vesicle Secretory Proteins; C424482:seminal vesicle-specific antigen; D006493:Heparin; D007781:Lactoferrin