Involvement of carbohydrate molecules on zona pellucida in human fertilization.
Although the involvement of several receptors and ligand molecules in sperm-zona interaction in many species have been proposed, there has been a little analysis of the kinetics between these molecules during the interaction. In the present study, we applied the detection method using surface plasmon resonance (SPR) by a BIAcore apparatus for the analysis of the putative receptor-ligand interaction of sperm-egg binding. Mannose-BSA or [man](5)-[GlcNAc](2)-Asp was immobilized on the surface of a sensor chip. When concanavalin A (Con A) was delivered to each of two different sensor chips to evaluate their usefulness, the resonance signal after sample injection onto a [man](5)-[GlcNAc](2)-Asp-fixed chip decreased rapidly than the mannose-BSA-fixed chip. However, the amount of binding for Con A during the injection onto the [man](5)-[GlcNAc](2)-Asp-fixed chip was high. When acid sperm extracts (acid extracts) and fractions through a CM column, containing protease activity (protease fractions), were delivered to the mannose-BSA-fixed chip, the SPR signal during the injection was not obviously changed compared with that of the control. However, when sperm samples were delivered to the [man](5)-[GlcNAc](2)-Asp-fixed chip, the SPR response during the injection was enormous. These results suggest that the [man](5)-[GlcNAc](2)-Asp-fixed chip is more useful than the mannose-BSA-fixed chip for investigating the interactions with sperm extracts and that the sensitive method using SPR by a BIAcore apparatus is applicable for the analysis of the putative receptor-ligand interaction of sperm-egg binding.
Maegawa
Masahiko
M
Department of Obstetrics and Gynecology, School of Medicine, The University of Tokushima, 3-18-15 Kuramoto-cho, Tokushima 770-8503, Japan. masahiko@clin.med.tokushima-u.ac.jp
Kamada
Masaharu
M
Yamamoto
Satoshi
S
Yamano
Shuji
S
Irahara
Minoru
M
Kido
Hiroshi
H
Aono
Toshihiro
T
D000818:Animals; D001224:Aspartic Acid / Q000031:analogs & derivatives; D002241:Carbohydrates / Q000737:chemistry / Q000502:physiology*; D002352:Carrier Proteins / Q000502:physiology; D002417:Cattle; D005260:Female; D005306:Fertilization / Q000502:physiology*; D006001:Glycoconjugates; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008358:Mannose; D037241:Mannose-Binding Lectins; D009844:Oligosaccharides; D012709:Serum Albumin; D013084:Sperm-Ovum Interactions / Q000502:physiology; D020349:Surface Plasmon Resonance; D015044:Zona Pellucida / Q000737:chemistry / Q000502:physiology*
D002241:Carbohydrates; D002352:Carrier Proteins; D006001:Glycoconjugates; D037241:Mannose-Binding Lectins; D009844:Oligosaccharides; D012709:Serum Albumin; C023840:mannose-bovine serum albumin conjugate; D001224:Aspartic Acid; D008358:Mannose