Structural requirements for incorporation of J chain into human IgM and IgA.
J chain is associated with pentameric IgM and dimeric IgA via disulfide bonds involving the penultimate cysteine residue in the secretory tailpiece of the mu or the alpha heavy chain. We have investigated the structural basis for incorporation of J chain by analyzing several IgM mutants, IgA mutants and IgG/IgM hybrid molecules. IgM mutants with the mu secretory tailpiece replaced by the alpha secretory tailpiece and/or Cys414 replaced by serine incorporated J chain, although in reduced amounts correlating with reduced pentamer/polymer formation. In addition to pentamers, tetramers of IgMC414S contained J chain, while no J chain was associated with smaller polymers or hexamers of IgM. An IgA/IgM hybrid tailpiece abolished J chain incorporation to pentameric IgM. Analysis of IgG molecules that have added a secretory tailpiece and/or have IgM domain replacements showed that J chain incorporation depends on regions of the C(mu)4 domain in addition to the tailpiece. Features of the C(mu)3 domain other than Cys414 also play a role in efficient formation of pentamers and J chain incorporation, while the C(mu)2 domain is not specifically required. By analysis of two IgA mutants that formed larger polymers than IgAwt, we found J chain equally incorporated into dimers, trimers, tetramers and pentamers. Thus, the results show that J chain incorporation into IgA does not depend on the polymeric structure, while J chain incorporation into IgM is restricted to certain polymeric conformations.
Sørensen
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Department of Molecular Cell Biology, Institute of Biology, University of Oslo, 0316 Oslo, Norway.
Rasmussen
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IB
Sundvold
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Michaelsen
T E
TE
Sandlie
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I
D006801:Humans; D007070:Immunoglobulin A / Q000235:genetics / Q000378:metabolism*; D007074:Immunoglobulin G / Q000378:metabolism; D007144:Immunoglobulin J-Chains / Q000378:metabolism*; D007075:Immunoglobulin M / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding
D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007144:Immunoglobulin J-Chains; D007075:Immunoglobulin M; C034639:polymeric IgA; C024242:polymeric IgM