title: Alternative pathways of disulfide bond formation yield secretion-competent, stable and functional immunoglobulins.
abstract: Disulfide bonds within and between proteins are responsible for stabilizing folding and covalent assembly. They are thought to form by an obligatory pathway that leads to a single native structure compatible with secretion. We have previously demonstrated that the intradomain disulfide in the C(H)1 domain of the Ig gamma2b heavy chains was dispensable for secretion [Elkabetz, Y., Argon, Y., Bar-Nun, S., 2005. Cysteines in C(H)1 underlie retention of unassembled Ig heavy chains. J. Biol. Chem. 280, 14402-14412]. Here we show that the heavy chain-light chain interchain disulfide is also dispensable. gamma2b with mutated Cys128, which normally disulfide bonds with the light chain, still assembled with lambdaI light chain into a secretion-competent, tetrameric IgG2b. This assembly comprised of a covalent homo-dimer of mutant heavy chains (C128S(2)) accompanied non-covalently by a covalent homo-dimer of light chains (lambda(2)). The lambda(2) homo-dimer formed only upon association with C128S(2), through disulfide bonding of the two "orphan" heavy chain-interacting Cys214 in lambdaI. The unique Ig tetramer was secreted efficiently as a functional antibody whose antigen-binding capacity resembled that of normal IgG2b. Therefore, disulfide bonding of Ig manifests considerable plasticity and can generate more than one functional structure that is considered native by the cellular quality control system.
authors.affiliation: Department of Biochemistry, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
authors.forename: Yechiel
authors.initials: Y
authors.lastname: Elkabetz
authors.forename: Ayala
authors.initials: A
authors.lastname: Ofir
authors.forename: Yair
authors.initials: Y
authors.lastname: Argon
authors.forename: Shoshana
authors.initials: S
authors.lastname: Bar-Nun
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003545:Cysteine; D004220:Disulfides / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000276:immunology; D007136:Immunoglobulins / Q000276:immunology*; D051379:Mice; D055503:Protein Multimerization; D055550:Protein Stability; D020836:Protein Structure, Quaternary
chemical_list: D004220:Disulfides; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D007136:Immunoglobulins; D003545:Cysteine