Determination of the type and quantity of sialic acid in the egg jelly coat of the sea urchin Paracentrotus lividus using capillary LC‐ESI‐MS/MS

Abstract

Sialic acid is a terminal sugar of carbohydrate chains that participates in numerous biological events. Recent studies have explored the mechanism of carbohydrate-mediated fertilisation to understand the biochemistry of fertilisation, although the type and quantity of sialic acid and the role of sialic acid during fertilisation remain unknown. Echinoderm fertilisation in particular has been studied extensively, yet our understanding of the mechanisms of carbohydrate-mediated fertilisation and the role of sialic acid remains incomplete. In this study, we characterised the sialic acid types in the egg jelly coat of the sea urchin, Paracentrotus lividus, using the sensitive analytical system capillary liquid chromatography electro-spray ionisation tandem mass spectrometry (capLC-ESI-MS/MS). First, we isolated the egg jelly coat and released its sialic acid using acid treatment. These sialic acids were derivatised with 1,2-diamino-4,5-methylenediaoxy-benzene dihydrochloride (DMB) and injected into the capLC-ESI-MS/MS system. When compared with standards, we identified twelve different types of sialic acid according to their retention times and collision-induced dissociation fragments. The mass spectral data revealed that Neu5Gc, Neu5Ac, Neu5GcS, and Neu5Gc9Ac were the predominant types of sialic acid in the sea urchin jelly coat, with Neu5Gc being the most abundant. Other types of sialic acid detected included Neu5AcS, Neu5Gc7,9Ac2, Neu5,9Ac2, Neu5Gc8Ac, Neu5Gc7Ac, Neu5,7Ac2, Neu5Gc8,9Ac2, and Neu5,8Ac2. The types and quantities of sialic acid that we detected in the egg jelly coat will aid in the discovery of new sialic acid-specific receptors on the sperm membrane.

Tilapia Lake Virus Does Not Hemagglutinate Avian and Piscine Erythrocytes and NH4Cl Does Not Inhibit Viral Replication In Vitro

“…Other viruses that use the Neu5,9Ac 2 receptor include coronaviruses. This receptor has been reported in various species, such as humans, rabbits, rats, guinea pigs, horses, chicken, turkey, goose, salmonids, crucian carp, mollusks, and sea urchins [40][41][42][43][44][45][46][47][48][49][50]. Unlike influenza C, ISAV specifically binds Neu4,5Ac 2 glycans on host cells using its hemagglutinin-esterase (HE) protein.…”

Profiling N-glycans of the egg jelly coat of the sea urchinParacentrotus lividusby MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems

“…Some hexoses are probably added on in the endoplamic reticulum membrane cytosolic zone, while the N-linked glycan biosynthetic pathway adds the other hexoses after the oligosaccharide precursor is flipped across the membrane into the lumen(de Leoz et al, 2011). The additional ionic glycans present on the jelly coat could be either sialic acids(Yeşilyurt et al, 2015) or other sulphated monosaccharides(Castro et al, 2009), although our analyzes suggest that no other substituted monosaccharides are present on these bulk chains.…”

“…Peptide N‐glycosidase F (PNGase‐F) treatment of the sea urchin egg jelly coat results in proteins of smaller relative mass, when compared with the non‐digested material, implying the presence of N ‐linked glycans on egg jelly glycoproteins (Keller & Vacquier, ). Sulfated glycans, polysaccharides, sulfated α‐L‐fucan, galactan, sialoglycoproteins, and acetylated sialic acids are present in the jelly coats of various sea urchin species, and each contributes a different, but essential, role during the sperm acrosome reaction and fertilization (Castro et al, ; Pomin, ; Valle et al, ; Vilela‐Silva, Alves, Valente, Vacquier, & Mourao, ; Yeşilyurt, Şahar, & Deveci, ). The importance of these glycan fragments in species recognition by gametes is further supported by the presence of novel, polymeric glycans, which are not commonly incorporated into eukaryotic glycoproteins.…”

“…Sulfated glycans, polysaccharides, sulfated α-L-fucan, galactan, sialoglycoproteins, and acetylated sialic acids are present in the jelly coats of various sea urchin species, and each contributes a different, but essential, role during the sperm acrosome reaction and fertilization (Castro et al, 2009;Pomin, 2015;Valle et al, 2015;Vilela-Silva, Alves, Valente, Vacquier, & Mourao, 1999;Yeşilyurt, Şahar, & Deveci, 2015).…”

Theory and Practice of UHPLC and UHPLC–MS

“…of the sea urchin (Yeşilyurt et al, 2015), serotonin, 5-hydroxytryptophan, and L-tryptophan in chocolate , or caffeine in tea, coffee, and cocoa (Al-Othman et al, 2012).…”