Purification and characterization of a surface lectin from the nematode-trapping fungus Arthrobotrys oligospora.
Several studies have indicated that the capture of nematodes by the nematophagous fungus Arthrobotrys oligospora is mediated by a lectin on the fungal surface. One of the major surface proteins of this fungus showed haemagglutinating activity and was isolated by affinity chromatography using a mucin Sepharose column. Biochemical analysis showed that the protein was a dimeric glycoprotein with a molecular mass of 36 kDa and an isoelectric point of pH 6.5, and contained no sulphur amino acids. The protein was N-terminally blocked; four internal peptides were sequenced, and showed no significant similarity to sequences in the Swiss-Prot or PIR databases. The haemagglutinating activity of the isolated protein was not inhibited by any of the mono- or disaccharides tested, but it was inhibited by the glycoproteins fetuin and mucin. The haemagglutinating activity changed after incubating the protein in buffers of different pH, with maximal activity at pH 11.0 and no activity at pH 2.8. The lectin was tested for different enzymic activities but none were detected. Analysis of the haemagglutinating activity in various cell fractions indicated that the protein was associated with extracellular polymer layers and with the cell wall of the fungus. About the same amount of the haemagglutinating protein was recovered from samples of vegetative mycelium and of mycelium containing nematode-trapping cells.
Rosén
S
S
Department of Microbial Ecology, Lund University, Sweden.
Ek
B
B
Rask
L
L
Tunlid
A
A
D000595:Amino Acid Sequence; D000818:Animals; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification*; D006031:Glycosylation; D006386:Hemagglutination Tests; D007526:Isoelectric Point; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*; D003904:Mitosporic Fungi / Q000737:chemistry*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009348:Nematoda / Q000382:microbiology
D005656:Fungal Proteins; D037102:Lectins; D008562:Membrane Glycoproteins