Contributions of a disulfide bond to the structure, stability, and dimerization of human IgG1 antibody CH3 domain.


Recombinant human monoclonal antibodies have become important protein-based therapeutics for the treatment of various diseases. The antibody structure is complex, consisting of beta-sheet rich domains stabilized by multiple disulfide bridges. The dimerization of the C(H)3 domain in the constant region of the heavy chain plays a pivotal role in the assembly of an antibody. This domain contains a single buried, highly conserved disulfide bond. This disulfide bond was not required for dimerization, since a recombinant human C(H)3 domain, even in the reduced state, existed as a dimer. Spectroscopic analyses showed that the secondary and tertiary structures of reduced and oxidized C(H)3 dimer were similar, but differences were observed. The reduced C(H)3 dimer was less stable than the oxidized form to denaturation by guanidinium chloride (GdmCl), pH, or heat. Equilibrium sedimentation revealed that the reduced dimer dissociated at lower GdmCl concentration than the oxidized form. This implies that the disulfide bond shifts the monomer-dimer equilibrium. Interestingly, the dimer-monomer dissociation transition occurred at lower GdmCl concentration than the unfolding transition. Thus, disulfide bond formation in the human C(H)3 domain is important for stability and dimerization. Here we show the importance of the role played by the disulfide bond and how it affects the stability and monomer-dimer equilibrium of the human C(H)3 domain. Hence, these results may have implications for the stability of the intact antibody. Study holds ProTherm entries: 23914, 23915, 23916, 23917 Extra Details: CH3 domain; reduced form human antibody; CH3 domain; disulfide bond; stability; dimerization

Submission Details

ID: ygmYZGs5

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:54 p.m.

Version: 1

Publication Details
McAuley A;Jacob J;Kolvenbach CG;Westland K;Lee HJ;Brych SR;Rehder D;Kleemann GR;Brems DN;Matsumura M,Protein Sci. (2008) Contributions of a disulfide bond to the structure, stability, and dimerization of human IgG1 antibody CH3 domain. PMID:18156469
Additional Information

Study Summary

Number of data points 4
Proteins Immunoglobulin heavy constant gamma 1
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: Cm ; Experimental Assay: Tm

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
199.4 H,K Immunoglobulin heavy constant gamma 1 P01857 IGHG1_HUMAN
181.2 H,K Immunoglobulin heavy constant gamma 1 P01859 IGHG2_HUMAN
181.2 H,K Immunoglobulin heavy constant gamma 1 P01861 IGHG4_HUMAN
198.2 L,M Immunoglobulin heavy constant gamma 1 P01834 IGKC_HUMAN