Amino acid sequence of the D-galactose binding lectin II from the sponge Axinella polypoides (Schmidt) and identification of the carbohydrate binding site in lectin II and related lectin I.
The sponge Axinella polypoides contains several D-galactose binding lectins. One of the main components, lectin I was sequenced earlier, the complete sequence of the other major constituent of saline extracts, lectin II has been determined by amino acid sequencing and mass spectrometry. Both lectins have a homology of 65% to each other and both possess a disulfide loop between positions 4 and 46. As long as this loop is closed in both lectins, they can be boiled in the presence of SDS or treated with 6 mol guanidine hydrochloride without losing their hemagglutinating activity. Incubation with beta-mercaptoethanol alone does not effect the carbohydrate binding capacity either. However, reduction of the disulfide bond under chaotropic conditions destroys the activity irreversibly. This disulfide loop is also an immunologically dominant epitope in both lectins, as was revealed with monospecific polyclonal antisera. Thus, sponge lectins seem to be of different origins, since three completely different structures were described: the structure of Geodia cydonium, related to the mammalian S-type lectins with one SH-group, the Axinella lectins with one disulfide loop and the Aaptos lectins I and II with 11 cysteine residues/subunit.
Buck
F
F
Institut für Zellbiologie und Klinische Neurobiologie, Hamburg, Germany.
Schulze
C
C
Breloer
M
M
Strupat
K
K
Bretting
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001665:Binding Sites / Q000235:genetics; D050260:Carbohydrate Metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D005690:Galactose / Q000378:metabolism*; D037161:Galectins; D006388:Hemagglutinins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D007120:Immunochemistry; D037102:Lectins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011161:Porifera / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid