Differential staining of Western blots of human secreted glycoproteins from serum, milk, saliva, and seminal fluid using lectins displaying diverse sugar specifities

Abstract

Human milk, serum, saliva, and seminal fluid glycoproteins (gps) nourish and protect newborn and adult tissues. Their saccharides, which resemble cell membrane components, may block pathogen adhesion and infection. In the present study, they were examined by a battery of lectins from plants, animals, and bacteria, using hemagglutination inhibition and Western blot analyses. The lectins included galactophilic ones from Aplysia gonad, Erythrina corallodendron, Maclura pomifera (MPL), peanut, and Pseudomonas aeruginosa (PA-IL); fucose-binding lectins from Pseudomonas aeruginosa (PA-IIL), Ralstonia solanacearum (RSL), and Ulex europaeus (UEA-I), and mannose/glucose-binding Con A. The results demonstrated the chosen lectin efficiency for differential analysis of human secreted gps as compared to CBB staining. They unveiled the diversity of these body fluid gp glycans (those of the milk and seminal fluid being highest): the milk gps interacted most strongly with PA-IIL, followed by RSL; the saliva gps with RSL, followed by PA-IIL and MPL; the serum gps with Con A and MPL, followed by PA-IIL and RSL, and the seminal plasma gps with RSL and MPL, followed by UEA-I and PA-IIL. The potential usage of these lectins as probes for scientific, industrial, and medical purposes, and for quality control of the desired gps is clearly indicated.

Lectin‐based glycoproteomics to explore and analyze hepatocellular carcinoma‐related glycoprotein markers

“…Briefly, after 2‐DE separation, total proteins are electrotransferred onto a PVDF membrane. Then glycoproteins are detected using peroxidase‐labeled lectins 50 and the corresponding glycoprotein profiles are obtained. Through comparing 2‐DE protein profiles with lectin‐blot glycoprotein profiles, glycosylated and/or aberrantly glycosylated proteins can be revealed.…”

Glycoproteins Presenting Galactose and N-Acetylgalactosamine in Human Seminal Plasma as Potential Players Involved in Immune Modulation in the Fertilization Process

“…MPL specificity has been described as GalNAc residue in the proximity of the peptide backbone, as Ser/Thr flanking amino acids may be bound by the lectin secondary binding site [ 59 ]. As elongation of the sugar chain with galactose does not change the binding, MPL is often reported as T-antigen specific [ 27 , 60 , 61 ]. Thus, the selected lectins enable the insight in glycans with the terminal, interactable residues of both monosaccharides of interest.…”