RRC ID 81871
著者 Yoshimoto S, Katayama K, Suzuki T, Dohmae N, Simizu S.
タイトル Regulation of N-glycosylation and secretion of Isthmin-1 by its C-mannosylation.
ジャーナル Biochim Biophys Acta Gen Subj
Abstract BACKGROUND:C-mannosylation is a type of protein glycosylation. Human Isthmin-1 (ISM1) is a 52-kDa secreted protein with a thrombospondin type 1 repeat (TSR) domain, containing two consensus C-mannosylation sequences at Trp223 and Trp226. In this study, we sought to examine the role of C-mannosylation in the secretion of ISM1.
METHODS:We established and cultured an ISM1-overexpressing HT1080 cell line and purified recombinant ISM1 for analysis from the conditioned medium by LC-MS/MS. Subcellular localization of ISM1 was observed by confocal fluorescence microscopy.
RESULTS:We found that ISM1 is C-mannosylated at Trp223 and Trp226 in the TSR domain. To determine the functions of the C-mannosylation of ISM1, we established a C-mannosylation-defective mutant ISM1-overexpressing HT1080 cell line and measured its secretion of ISM1. The secretion of ISM1 decreased significantly in this mutant ISM1-overexpressing line compared with wild-type cells. Furthermore, ISM1 was N-glycosylated only in these C-mannosylation-defective cells.
CONCLUSIONS:ISM1 is C-mannosylated in its TSR domain, and the status of the C-mannosylation of ISM1 affects its N-glycosylation.
GENERAL SIGNIFICANCE:The C-mannosylation of ISM1 regulates its N-glycosylation status.
巻・号 1865(3)
ページ 129840
公開日 2021-3-1
DOI 10.1016/j.bbagen.2020.129840
PII S0304-4165(20)30351-2
PMID 33412225
MeSH Amino Acid Motifs Amino Acid Substitution Cell Line, Tumor Culture Media, Conditioned / chemistry Fibroblasts / cytology Fibroblasts / metabolism Gene Expression Genetic Vectors / chemistry Genetic Vectors / metabolism Glycosylation Humans Mannose / chemistry Mannose / metabolism* Mutation Protein Processing, Post-Translational* Recombinant Proteins / genetics Recombinant Proteins / metabolism Thrombospondins / genetics Thrombospondins / metabolism*
IF 3.422
リソース情報
遺伝子材料 ISM1-MH (RDB18696) ISM1 2WF-MH (RDB18697) ISM1 (N39Q) (RDB18698) ISM1 (N285Q) (RDB18699) ISM1 (N39Q/N285Q) (RDB18700) ISM1/2WF (N39Q) (RDB18701) ISM1/2WF (N285Q) (RDB18702) ISM1/2WF(N39Q/N285Q) (RDB18703)