A novel recombinantly produced banana lectin isoform is a valuable tool for glycoproteomics and a potent modulator of the proliferation response in CD3+, CD4+, and CD8+ populations of human PBMCs.
Lectins as carbohydrate-binding proteins have been employed in various biological assays for the detection and characterization of glycan structures on glycoproteins, including clinical biomarkers in disease states. A mannose-specific banana lectin (BanLec) is unique in its specificity for internal alpha1,3 linkages as well as beta1,3 linkages at the reducing termini. The immunomodulatory potential of natural BanLec was recognized by a strong immunoglobulin G4 antibody response and T cell mitogen activity in humans. To explore its applicability in glycoproteomics and its modulatory potential, the gene of banana lectin was cloned, sequenced and a recombinant protein was produced in Escherichia coli. The obtained cDNA revealed a novel banana lectin isoform, with an open reading frame of 426 nucleotides, encoding a cytoplasmatic protein of 141 amino acids. The molecular mass of rBanLec determined by ESI FT-MS and N-terminal sequencing confirmed the cDNA at the protein level. The specificity of rBanLec for detection glycan structures was the same as for natural BanLec as examined with five protein extracts rich in glycoprotein content, as well as with horseradish peroxidase glycoprotein. Besides, the immunomodulatory potential of rBanLec and nBanLec were comparable as assessed by an inhibition assay and a human T cell proliferation assay where they induced a strong proliferation response in CD3+, CD4+, and CD8+ populations of human PBMCs. This recombinant BanLec is a useful reagent for glycoproteomics and lectin microarrays, with a potential for modulation of the immune response.
Gavrovic-Jankulovic
Marija
M
Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Belgrade, Serbia. mgavrov@chem.bg.ac.yu
Poulsen
Knud
K
Brckalo
Tamara
T
Bobic
Sonja
S
Lindner
Buko
B
Petersen
Arnd
A
D000595:Amino Acid Sequence; D017252:CD3 Complex / Q000032:analysis; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D054794:Glycomics; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007155:Immunologic Factors / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D008358:Mannose / Q000032:analysis*; D037241:Mannose-Binding Lectins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000737:chemistry; D028521:Musa / Q000737:chemistry*; D037121:Plant Lectins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D040901:Proteomics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D016415:Sequence Alignment
D017252:CD3 Complex; D006023:Glycoproteins; D007074:Immunoglobulin G; D007155:Immunologic Factors; D037241:Mannose-Binding Lectins; D009005:Monosaccharides; D037121:Plant Lectins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D008358:Mannose