Sequence tolerance of immunoglobulin variable domain framework regions to noncanonical intradomain disulfide linkages.

ABSTRACT: Most immunoglobulin (Ig) domains bear only a single highly conserved canonical intradomain, inter-β-sheet disulfide linkage formed between Cys23-Cys104, and incorporation of rare noncanonical disulfide linkages at other locations can enhance Ig domain stability. Here, we exhaustively surveyed the sequence tolerance of Ig variable (V) domain framework regions (FRs) to noncanonical disulfide linkages. Starting from a destabilized V_H domain lacking a Cys23-Cys104 disulfide linkage, we generated and screened phage-displayed libraries of engineered V_Hs, bearing all possible pairwise combinations of Cys residues in neighboring β-strands of the Ig fold FRs. This approach identified seven novel Cys pairs in V_H FRs (Cys4-Cys25, Cys4-Cys118, Cys5-Cys120, Cys6-Cys119,

SUBMITTER: Kim DY 

PROVIDER: S-EPMC10641266 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature