Reassembly of Immunoglobulin M Heavy and Light Chains In Vitro
Reassembly of Immunoglobulin M Heavy and Light Chains In Vitro
复制标题
免疫球蛋白 M 重链和轻链的体外重组
DOI：
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发表时间：
1976
影响因子：
3.7
通讯作者：
R. Hester
中科院分区：
文献类型：
--
作者：
R. Schrohenloher;R. Hester
Reduced and alleviated monoclonal IgM was fractionated into μ and light (L) chains by gel chromatography in 1N acetic acid. Equimolar mixtures of the chains formed a noncovalently bonded structure in 0.01M sodium acetate buffer, pH 4.1, that had the properties of a half subunit. The latter reassociated into a subunit‐like structure after transfer into 0.08M sodium phosphate buffer, pH 7.5. The similarity of the reconstituted IgM subunit (IgMs) to that of the native molecule was established by its physicochemical and immunochemical properties. Comparable products were obtained on reassembly of the alkylated μ and L chains from several other monoclonal IgM. The presence of active binding sites for IgG on subunits reconstituted from the chains of proteins with anti‐IgG activity further indicated correct assembly of the μ and L chains. High yields of subunit‐like products were also obtained by assembly of μ chains from one protein and L chains from another. Evidence was obtained that L chains of appropriate specificity can substitute for the homologous chain in the formation of the active site. Heterogeneous mixtures of high molecular weight products were generated from μ and L chains that were not alkylated. Reduction and alkylation demonstrated that the products represented polymers of reconstituted IgMs. Significant levels of anti‐IgG activity were detected in the polymeric IgM generated from the chains of active proteins by precipitation with aggregated IgG.
