Isolation and characterization of J-chain from bovine colostral immunoglobulin M.
Purified bovine colostral intact immunoglobulin M (IgM) exhibited the presence of an anodal, single, fast moving band (noncovalently bound form) when subjected to analytical polyacrylamide gel electrophoresis at an alkaline pH in urea. Reduced and alkylated or sulfitolysed bovine colostral IgM (devoid of the noncovalently bound form) also showed the presence of a similar band (covalently bound form). The molecular weight of both the covalently bound and noncavalently bound forms of the fast component was determined to be 16,500 by sodium dodecyl sulfate - polyacrylamide gel electrophoresis. In addition, the non-cavalently bound form of the fast-moving component was found to be antigenically identical to the covalently bound form. The noncovalently bound form sedimented as a single peak at 1.56 S. Antiserum against the fast-moving component precipitated neither bovine colostral IgG nor mu-chains and bovine serum albumin, but precipitated native or denatured intact IgM (devoid of the non-covalently bound form) and human J-chains and vice versa, thus permitting the fast-moving components to be classified as J-chains. Radioalkylation experiments revealed the presence of 9.7 sulfhydryl groups per mole, for both the covalently and non-covalently bound forms of bovine J-chain. The stoichiometry of J-chain, determined from the densitometric tracing of the reduced and alkylated bovine colostral IgM (devoid of the noncovalently bound J-chain) in stained analytical polyacrylamide gels, revealed the presence of one J-chain per IgM molecule. On the other hand the amount of non-covalently bound form of J-chain was determined to be 1.2 per molecule of IgM.
D000818:Animals; D002417:Cattle; D003126:Colostrum / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007144:Immunoglobulin J-Chains / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis*; D008970:Molecular Weight; D011247:Pregnancy
D007144:Immunoglobulin J-Chains; D007075:Immunoglobulin M