Involvement of glycans in the immunological cross-reaction between alpha-macroglobulin and hemocyanin of the gastropod Helix pomatia.
By tandem-crossed immunoelectrophoresis and ELISA experiments an immunological relationship was observed between alpha-macroglobulin (alphaM) and hemocyanin (Hc) of the terrestrial snail Helix pomatia. Both glycoproteins occur in the hemolymph: alphaM (minor component) as a specific proteinase inhibitor, Hc (consisting of three components: alpha(D)-HpH, alpha(N)-HpH and beta-HpH) as oxygen transport protein. The cross-reaction was found to be correlated with glycosylation. (i) With beta-HpH, which is richer in carbohydrates than alpha(D)-HpH and alpha(N)-HpH, mainly due to a higher 3-O-methyl-d-galactose content, the cross-reaction with HpalphaM was highest. (ii) From the 8 functional units, designated a-h, isolated from beta-HpH, two that lack carbohydrates (c and f) were not recognized by antibodies against HpalphaM, while the six glycosylated ones were strongly cross-reacting. The nearly complete loss of the cross-reactivity upon deglycosylation of functional units d and g and the inhibition in competitive ELISA experiments by glycopeptides isolated from both beta-HpH and HpalphaM are further evidence that glycans are involved in the immunological relationship between HpH and HpalphaM. Carbohydrate analyses indicated that the glycan structures present on HpalphaM are very similar (or identical) to those found on HpH, suggesting that glycans are common epitopes on both proteins. Especially d-xylose and 3-O-methyl-d-galactose seem to be responsible for the cross-reactivity since the alpha-macroglobulin and hemocyanin of the cephalopod Sepia officinalis, which lack these two monosaccharides in their glycan structures, do not immunologically cross-react.
Siddiqui
Nurul I
NI
Division of Biochemistry, Molecular and Structural Biology, Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200G, B-3001 Leuven-Heverlee, Belgium.
Yigzaw
Yinges
Y
Préaux
Gisèle
G
Gielens
Constant
C
D000818:Animals; D003429:Cross Reactions / Q000276:immunology; D006372:Helix, Snails / Q000276:immunology*; D006433:Hemocyanins / Q000276:immunology* / Q000302:isolation & purification; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D000511:alpha-Macroglobulins / Q000276:immunology* / Q000302:isolation & purification
D011134:Polysaccharides; D000511:alpha-Macroglobulins; D006433:Hemocyanins