New GlcNAc/GalNAc-specific lectin from the ascidian Didemnum ternatanum.
Previously we isolated GlcNAc-specific lectin (DTL) from the ascidian Didemnum ternatanum by affinity chromatography on cross-linked ovalbumin. Here we report the purification and characterization of new D-GlcNAc/D-GalNAc-specific lectin DTL-A from the same ascidian. This lectin was isolated from non-bound cross-linked ovalbumin fraction and further was purified by gel filtration on Sepharose CL-4B, affinity chromatography on GlcNAc-agarose and gel filtration on Superdex 200. SDS-polyacrylamide gel electrophoresis and gel filtration of purified lectin on Sepharose CL-4B indicates that it exists as large aggregates in the native state. Investigations of the carbohydrate specificity of DTL-A by enzyme-linked lectin assay suggest the multi-specificity of this lectin. DTL-A binds BSM, asialo-BSM as well as heparin and dextran sulfate. The binding of DTL-A to BSM was inhibited by monosaccharides D-GlcNAc and D-GalNAc, their alpha- but not beta-anomers. Among polysaccharides and glycoconjugates, DTL-A binding to BSM was effectively inhibited by BSM, asialo-BSM, pronase-treated BSM and synthetic alpha-D-GalNAc-PAA. Fetuin and asialofetuin showed a much lower inhibitory potency, heparin and dextran sulfate were noninhibitory. On the other hand, DTL-A binding to heparin was effectively inhibited by dextran sulfate, fucoidan, whereas BSM showed insignificantly inhibitory effect. DTL-A binding to heparin was not inhibited by D-GlcNAc and D-GalNAc.
Molchanova
Valentina
V
Pacific Institute of Bioorganic Chemistry, Far East, Branch of the Russian Academy of Science, Prospect 100-letiya Vladivostoku, 159, 690022, Vladivostok, Russia.
Chikalovets
Irina
I
Li
Wei
W
Kobelev
Stanislav
S
Kozyrevskaya
Svetlana
S
Bogdanovich
Raisa
R
Howard
Eric
E
Belogortseva
Natalia
N
D000116:Acetylgalactosamine / Q000378:metabolism*; D000117:Acetylglucosamine / Q000378:metabolism*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002846:Chromatography, Affinity; D006384:Hemagglutination; D006493:Heparin / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D009077:Mucins / Q000378:metabolism; D009994:Osmolar Concentration; D014561:Urochordata / Q000737:chemistry*
D000596:Amino Acids; C115440:BSM2 protein, Bos taurus; D037102:Lectins; D009077:Mucins; D006493:Heparin; D000116:Acetylgalactosamine; D000117:Acetylglucosamine