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

Int J Oncol. 2002 Feb;20(2):367-72.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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

Substances

  • Biomarkers, Tumor
  • Glycoproteins
  • Lectins
  • Membranes, Artificial
  • Concanavalin A
  • Fucose
  • N-Acetylneuraminic Acid