Lectin isolation and detection of N-glycoproteins bearing sialic acid and L-fucose residues in human colorectal mucosa and in adenocarcinoma biopsies

• Published in: International journal of oncology Vol. 20; no. 2; p. 367
• Main Authors: De Carlos, Alejandro, Caride-Castro, Amado, Martinez-Zorzano, Vicenta S, De La Cadena, María Páez, Rodríguez-Berrocal, Francisco J
• Format: Journal Article
• Language: English
• Published: Greece 01.02.2002
• Subjects: Adenocarcinoma - chemistry; Biomarkers, Tumor - analysis; Biomarkers, Tumor - chemistry; Biopsy; Chromatography, Affinity; Chromatography, High Pressure Liquid; Colorectal Neoplasms - chemistry; Concanavalin A - metabolism; Electrophoresis, Polyacrylamide Gel; Fucose - analysis; Glycoproteins - analysis; Glycoproteins - chemistry; Humans; Intestinal Mucosa - chemistry; Lectins - isolation & purification; Lectins - metabolism; Membranes, Artificial; Molecular Weight; N-Acetylneuraminic Acid - analysis; Organ Specificity
• ISSN: 1019-6439
• DOI: 10.3892/ijo.20.2.367

A method to improve the reactivity to specific lectins of N-glycoproteins isolated from human colorectal mucosa and from adenocarcinoma biopsies was developed using a combination of techniques. Total protein extracts were subjected to affinity chromatography, using the immobilised lectin Concanavalin A coupled to Sepharose, by fast performance liquid chromatography (FPLC). N-glycoprotein enriched fractions were resolved by SDS-PAGE, transferred to PVDF membranes and incubated with various lectins. Digoxigenin-conjugated SNA I and MAA lectins were used to detect sialic acid residues. Biotin-conjugated UEA I lectin was used to detect L-fucose residues. By this method, lectin-binding N-glycoproteins were found in a broad relative molecular mass (Mr) range (from 47 to 205 kDa). No tissue-specific N-glycoproteins were observed when human colorectal mucosa and adenocarcinoma samples were compared.