Effect of O-glycosylation on amyloid fibril formation of the variable domain in the Vλ6 light chain mutant Wil

• Published in: International journal of biological macromolecules 2021-01, Vol.166, p.342-351
• Main Authors: Abe, Yoshito, Shibata, Hinako, Oyama, Kousuke, Ueda, Tadashi
• Format: Article
• Language: English
• Subjects: AL amyloidosis; Amyloid - chemistry; Amyloid - genetics; Amyloid - metabolism; Amyloid fibril formation; Glycosylation; Humans; Immunoglobulin lambda-Chains - chemistry; Immunoglobulin lambda-Chains - genetics; Immunoglobulin lambda-Chains - metabolism; Immunoglobulin Variable Region - chemistry; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - metabolism; Mutation; O-glycosylation; Protein Multimerization; Protein Processing, Post-Translational; Protein Stability; Saccharomycetales
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2020.10.194

Glycosylation is one of the major post-translational modifications in eukaryotic cells and has been reported to affect the amyloid fibril formation in several amyloidogenic proteins and peptides. In this study, we expressed a Vλ6 light chain mutant, Wil, which is an amyloidogenic mutant in AL amyloidosis, by the yeast Pichia pastoris. After separation by cation exchange chromatography, we obtained the O-glycosylated and non-glycosylated Wil mutants in high yield. The structures of these Wil mutants were identical except with respect to glycosylation, and the stabilities were also identical. On the other hand, the O-glycosylation retarded the amyloid fibril formation in a sugar size-dependent manner. From these results, we discussed the role of covalently attached glycan in the retardation of amyloid fibril formation. •Wil mutants with O-glycans are expressed in yeast Pichia Pastris and are purified.•Glycosylated and non-glycosylated Wil mutants have same structures and stabilities.•Retardation of amyloid fibril formation depends on a sugar size in Wil mutant.