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Horm Metab Res 2011; 43(08): 545-550
DOI: 10.1055/s-0031-1280782
Original Basic
Georg Thieme Verlag KG Stuttgart · NewYork

N-Linked Glycosylation of Mouse Adiponectin

M. Tanaka
1   Department of Metabolic Medicine, Graduate School of Medicine, Osaka University, Osaka, Japan
,
A. Fukuhara
1   Department of Metabolic Medicine, Graduate School of Medicine, Osaka University, Osaka, Japan
,
I. Shimomura
1   Department of Metabolic Medicine, Graduate School of Medicine, Osaka University, Osaka, Japan
› Author Affiliations
Further Information

Publication History

received 13 September 2010

accepted 25 May 2011

Publication Date:
18 July 2011 (online)

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Abstract

Adiponectin is an insulin-sensitizing adipokine with antidiabetic, anti-atherogenic, anti-inflammatory, and cardioprotective properties. Previously, some types of posttranslational modification on adiponectin have been reported. In this study, we demonstrate that mouse adiponectin protein migrated as 2 bands on SDS-PAGE gel. Slower migrating band of adiponectin was reduced by PNGase treatment. PNGase is known as N-glycosidase, and is able to change the mobility of N-glycosylated protein on SDS-PAGE gel. This result indicates the possibility that slower band shifted and overlapped with faster band by cleavage of N-glycan. To further clarify the N-glycosylation of adiponectin, we investigated the effect of N-glycosylation inhibitor tunicamycin on 3T3-L1 adipocytes. Tunicamycin significantly reduced the ratio of slower band to faster band in culture medium from 3T3-L1 adipocytes. This result also indicates the possibility that slower band of adiponectin is N-glycosylated. Lastly, to identify glycosylated asparagine residues, we established 3T3-L1 cell lines stably expressing wild type and mutant adiponectin in N-glycosylation sites. Wild-type adiponectin protein migrated as double bands, and mutant adiponectin in either asparagine at position 53 or threonine at 55 lacked slower band. These results suggest that a part of mouse adiponectin is modified by N-linked glycosylation at asparagine 53.

Key words

adiponectin - modification - adipocyte
 

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