Extensive Crosstalk between O-GlcNAcylation and Phosphorylation Regulates Akt Signaling

被引：93

作者：

Wang, Shuai ^{[1
]}

Huang, Xun ^{[1
]}

Sun, Danni ^{[1
]}

Xin, Xianliang ^{[1
]}

Pan, Qiuming ^{[1
]}

Peng, Shuying ^{[2
]}

Liang, Zhongjie ^{[3
]}

Luo, Cheng ^{[3
]}

Yang, Yiming ^{[2
]}

Jiang, Hualiang ^{[3
]}

Huang, Min ^{[1
]}

Chai, Wengang ^{[1
,4
]}

Ding, Jian ^{[1
]}

Geng, Meiyu ^{[1
]}

机构：

[1] Chinese Acad Sci, Shanghai Inst Mat Med, Div Antitumor Pharmacol, Shanghai 200031, Peoples R China

[2] Chinese Acad Sci, Shanghai Inst Mat Med, Lab Mass Spectrometry, Shanghai 200031, Peoples R China

[3] Chinese Acad Sci, Shanghai Inst Mat Med, Ctr Drug Discovery & Design, State Key Lab Drug Res, Shanghai 200031, Peoples R China

[4] Imperial Coll London, Glycosci Lab, Fac Med, Harrow, Middx, England

来源：

PLOS ONE | 2012年 / 7卷 / 05期

关键词：

BETA-N-ACETYLGLUCOSAMINE; PROTEIN-KINASE B; INSULIN-RESISTANCE; IN-VIVO; GLCNAC; IDENTIFICATION; ACTIVATION; DYNAMICS; GROWTH; SITE;

D O I：

10.1371/journal.pone.0037427

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

O-linked N-acetylglucosamine glycosylations (O-GlcNAc) and O-linked phosphorylations (O-phosphate), as two important types of post-translational modifications, often occur on the same protein and bear a reciprocal relationship. In addition to the well documented phosphorylations that control Akt activity, Akt also undergoes O-GlcNAcylation, but the interplay between these two modifications and the biological significance remain unclear, largely due to the technique challenges. Here, we applied a two-step analytic approach composed of the O-GlcNAc immunoenrichment and subsequent O-phosphate immunodetection. Such an easy method enabled us to visualize endogenous glycosylated and phosphorylated Akt subpopulations in parallel and observed the inhibitory effect of Akt O-GlcNAcylations on its phosphorylation. Further studies utilizing mass spectrometry and mutagenesis approaches showed that O-GlcNAcylations at Thr 305 and Thr 312 inhibited Akt phosphorylation at Thr 308 via disrupting the interaction between Akt and PDK1. The impaired Akt activation in turn resulted in the compromised biological functions of Akt, as evidenced by suppressed cell proliferation and migration capabilities. Together, this study revealed an extensive crosstalk between O-GlcNAcylations and phosphorylations of Akt and demonstrated O-GlcNAcylation as a new regulatory modification for Akt signaling.

引用

页数：11

共 47 条

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