Partial characterisation of high-molecular weight glycoconjugates in the trail mucus of the freshwater pond snail Lymnaea stagnalis.
We have studied the glycoconjugates in trail mucus of the pond snail Lymnaea stagnalis. The mucus was dissolved with 6 M guanidinium hydrochloride (GuHCl) and the major component was comprised of very high-M(r) glycoconjugates that were eluted in the void volume of a Sepharose CL-4B gel-filtration column. This high-M(r) material was pooled and thereafter subjected to density gradient centrifugation first in 4 M GuHCl/CsCl and subsequently 0.2 M GuHCl/CsCl to further remove non-glycosylated proteins and DNA. The harvested glycoconjugate pool chromatographed in the void volume of Sepharose CL-2B. However, reduction of disulfide bonds lowered the molecular size of approximately 80% of the void material yielding a major fragment and some minor smaller fragments in gel chromatography. The reduced glycoconjugates were digested with papain and yielded high molecular weight, proteinase-resistant glycopeptides. This fragmentation pattern is similar to that found for oligomeric gel-forming mucins in mammals and the amino acid composition (60% Ser/Thr) and sugar analysis of the glycopeptides is consistent with mucin-like molecules, there being no significant amounts of xylose or uronic acids. The residual 20% of the preparation, which apparently resisted reduction and protease digestion, had a similar amino acid composition to the bulk, but was somewhat different in sugar composition, containing some xylose and a significant amount of glucuronic acid. The two groups of molecules had very different morphologies in the electron microscope. Taken together, these data suggest that trail mucus is a complex mixture of at least two families of protein-glycoconjugate molecules based upon the gel-forming mucin and proteoglycan families, though we cannot rule out that polysaccharides may also be present.
Ballance
Simon
S
School of Biological Sciences, University of Manchester, Manchester M13 9PL, UK. simon.ballance@biotech.ntnu.no
Howard
Marj
M
White
Keith N
KN
McCrohan
Catherine R
CR
Thornton
David J
DJ
Sheehan
John K
JK
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002241:Carbohydrates / Q000737:chemistry; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D005618:Fresh Water; D020723:Glucuronic Acid / Q000737:chemistry; D006001:Glycoconjugates / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006020:Glycopeptides / Q000737:chemistry; D008195:Lymnaea / Q000737:chemistry* / Q000378:metabolism; D008970:Molecular Weight; D009093:Mucus / Q000737:chemistry* / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D013329:Structure-Activity Relationship; D014994:Xylose / Q000737:chemistry
D000596:Amino Acids; D002241:Carbohydrates; D006001:Glycoconjugates; D006020:Glycopeptides; D020723:Glucuronic Acid; D014994:Xylose; D010447:Peptide Hydrolases