522 338

Cited 12 times in

N-linked glycosylation plays a crucial role in the secretion of HMGB1

Authors
 Young Hun Kim  ;  Man Sup Kwak  ;  Jun Bae Park  ;  Shin-Ae Lee  ;  Ji Eun Choi  ;  Hyun-Soo Cho  ;  Jeon-Soo Shin 
Citation
 JOURNAL OF CELL SCIENCE, Vol.129(1) : 29-38, 2016 
Journal Title
 JOURNAL OF CELL SCIENCE 
ISSN
 0021-9533 
Issue Date
2016
MeSH
Amino Acid Sequence ; Animals ; CHO Cells ; Cell Nucleus/metabolism ; Chromatography, Liquid ; Cricetinae ; Cricetulus ; DNA/metabolism ; Glycosylation ; HEK293 Cells ; HMGB1 Protein/chemistry ; HMGB1 Protein/secretion* ; HeLa Cells ; Humans ; Intracellular Space/metabolism ; Karyopherins/metabolism ; Mice ; Molecular Sequence Data ; Mutant Proteins/metabolism ; Polysaccharides/chemistry ; Polysaccharides/metabolism ; Protein Binding ; Protein Stability ; Protein Transport ; Receptors, Cytoplasmic and Nuclear/metabolism ; Tandem Mass Spectrometry
Keywords
DNA binding ; HMGB1 ; N-glycosylation ; Post-translational modification ; Secretion
Abstract
HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. Here, we identified the role of N-glycosylation of HMGB1 in extracellular secretion. We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. Inhibition of N-glycosylation with tunicamycin resulted in a molecular shift of HMGB1 as assessed by gel electrophoresis. Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. These mutant proteins had reduced secretion even after acetylation, phosphorylation, oxidation and exposure to pro-inflammatory stimuli. Taken together, we propose that HMGB1 is N-glycosylated, and that this is important for its DNA interaction and is a prerequisite for its nucleocytoplasmic transport and extracellular secretion.
Files in This Item:
T201600218.pdf Download
DOI
10.1242/jcs.176412
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Kwak, Man Sup(곽만섭) ORCID logo https://orcid.org/0000-0002-3989-3016
Shin, Jeon Soo(신전수) ORCID logo https://orcid.org/0000-0002-8294-3234
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/146317
사서에게 알리기
  feedback

qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse