Identification, purification and properties of a beta-1,3-glucan-specific lectin from the serum of the cockroach, Blaberus discoidalis which is implicated in immune defence reactions.
A lectin specific for laminarin, a beta-1,3-glucan, agglutinating baker's yeast and enhancing prophenoloxidase activation by laminarin, has been purified from the cockroach, Blaberus discoidalis, serum. Purification involved gel filtration with Bio-gel P300 and affinity chromatography on blue Sepharose CL-6B and laminarin-Sepharose 4B. The purified lectin has a molecular mass estimate of 520 kDa determined by gel filtration, and approximately 80 and 82 kDa by SDS-PAGE, under non-reducing and reducing conditions, respectively. After isoelectric focusing the lectin focused as a single band at pH 4.9. The purified lectin was stained by the periodic acid/Schiff's reagent showing that it is a glycoprotein, and was deglycosylated by endo-beta-N-acetylglu-cosaminidase F. Amino acid composition analysis showed the protein is similar to previously purified beta-1,3-glucan binding proteins from other invertebrates. In electron micrographs by negative staining, the protein formed large aggregates with 'Y'-shaped 'structural units' ca. 79 x 65 nm. Immunological tests confirmed that this lectin is not related to any other lectins previously purified from the same insect. This protein appears to be part of the hexamerin family of proteins. This is one of the first reports of a hexamerin-like molecule with lectin activity.
Chen
C
C
Biomedical and Physiological Research Group, School of Biological Sciences, University of Wales Swansea, Wales, UK.
Rowley
A F
AF
Newton
R P
RP
Ratcliffe
N A
NA
D000372:Agglutination Tests; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D004156:Catechol Oxidase / Q000378:metabolism; D003058:Cockroaches / Q000276:immunology*; D004792:Enzyme Precursors / Q000378:metabolism; D005260:Female; D005936:Glucans / Q000378:metabolism*; D006023:Glycoproteins / Q000737:chemistry; D006031:Glycosylation; D006433:Hemocyanins / Q000737:chemistry; D006434:Hemocytes / Q000737:chemistry; D006801:Humans; D007106:Immune Sera; D019476:Insect Proteins / Q000737:chemistry; D037102:Lectins / Q000276:immunology* / Q000302:isolation & purification* / Q000378:metabolism; D008297:Male; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D047071:beta-Glucans
D000596:Amino Acids; D002241:Carbohydrates; D004792:Enzyme Precursors; D005936:Glucans; D006023:Glycoproteins; D007106:Immune Sera; D019476:Insect Proteins; D037102:Lectins; D010446:Peptide Fragments; D047071:beta-Glucans; C041137:storage proteins, Insecta; D006433:Hemocyanins; C033363:beta-1,3-glucan; C038451:pro-phenoloxidase; D004156:Catechol Oxidase