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Structural and functional analysis of J chain-deficient IgM.

Journal of immunology (Baltimore, Md. : 1950) (1998-06-24)
E J Wiersma, C Collins, S Fazel, M J Shulman
RESUMEN

Previous studies have discerned two forms of polymeric mouse IgM: moderately cytolytic (complement-activating) pentamer, which contains J chain, and highly cytolytic hexamer, which lacks J chain. To investigate the relationships among polymeric structure, J chain content, and cytolytic activity, we produced IgM in J chain-deficient and J chain-proficient mouse hybridoma cell lines. Both hexamer and pentamer were produced in the absence as well as the presence of J chain. Hexameric IgM activated (guinea pig) complement approximately 100-fold more efficiently than did J chain-deficient pentamer, which, in turn, was more active than J chain-containing pentamer. These results are consistent with the hypothesis that J chain-containing pentamer cannot activate complement. We also analyzed the structure of IgM-S337, in which the mu-chain bears the C337S substitution. Like normal IgM, IgM-S337 was formed as a hexamer and as both J chain deficient- and J chain-containing pentamers. Unlike normal IgM, IgM-S337 dissociated in SDS into various subunits. For IgM-S337 pentamer, the predominant subunits migrated as mu2kappa2 and mu4kappa4, and the subunit distribution was unaltered by J chain. However, J chain was found only in the mu2kappa2 species, suggesting that some arrangement of inter-mu bonds directs incorporation of J chain. IgM-S337 hexamer also dissociated to mu2kappa2 and mu4kappa4, but also yielded several species migrating much more slowly in SDS-PAGE than wild-type mu12kappa12. To account for these forms, we propose that each mu-chain can interact with three other mu-chains and that some hexameric molecules contain two catenated mu6kappa6 circles.

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