Mapping and Quantification of Over 2000 O-linked Glycopeptides in Activated Human T Cells with Isotope-Targeted Glycoproteomics (Isotag).

糖肽 聚糖 N-糖酰胺酶F 糖组学 质谱法 色谱法 生物化学 分子生物学 串联质谱法
作者
Christina M. Woo,Peder Lund,Andrew C. Huang,Mark M. Davis,Carolyn R. Bertozzi,Sharon J. Pitteri
出处
期刊:Molecular & Cellular Proteomics [Elsevier BV]
卷期号:17 (4): 764-775 被引量:96
标识
DOI:10.1074/mcp.ra117.000261
摘要

Post-translational modifications (PTMs) on proteins often function to regulate signaling cascades, with the activation of T cells during an adaptive immune response being a classic example. Mounting evidence indicates that the modification of proteins by O-linked N-acetylglucosamine (O-Glcnac), the only mammalian glycan found on nuclear and cytoplasmic proteins, helps regulate T cell activation. Yet, a mechanistic understanding of how O-Glcnac functions in T cell activation remains elusive, partly because of the difficulties in mapping and quantifying O-Glcnac sites. Thus, to advance insight into the role of O-Glcnac in T cell activation, we performed glycosite mapping studies via direct glycopeptide measurement on resting and activated primary human T cells with a technique termed Isotope Targeted Glycoproteomics. This approach led to the identification of 2219 intact O-linked glycopeptides across 1045 glycoproteins. A significant proportion (>45%) of the identified O-Glcnac sites lie near or coincide with a known phosphorylation site, supporting the potential for PTM crosstalk. Consistent with other studies, we find that O-Glcnac sites in T cells lack a strict consensus sequence. To validate our results, we employed gel shift assays based on conjugating mass tags to O-Glcnac groups. Notably, we observed that the transcription factors c-JUN and JUNB show higher levels of O-Glcnac glycosylation and higher levels of expression in activated T cells. Overall, our findings provide a quantitative characterization of O-Glcnac glycoproteins and their corresponding modification sites in primary human T cells, which will facilitate mechanistic studies into the function of O-Glcnac in T cell activation.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
王子安应助Stardust采纳,获得10
1秒前
充电宝应助顺利涵菡采纳,获得10
3秒前
4秒前
Buduan完成签到,获得积分10
5秒前
5秒前
Ma关注了科研通微信公众号
7秒前
7秒前
8秒前
畅快城发布了新的文献求助10
8秒前
复成完成签到 ,获得积分10
9秒前
yinle关注了科研通微信公众号
9秒前
aktuell完成签到,获得积分10
9秒前
9秒前
ANG发布了新的文献求助10
9秒前
Lucas应助2116564采纳,获得10
9秒前
量子星尘发布了新的文献求助10
10秒前
清辉夜凝发布了新的文献求助10
10秒前
可爱的函函应助Shrine采纳,获得10
11秒前
recovery应助Aprilapple采纳,获得10
12秒前
baonali发布了新的文献求助10
13秒前
123发布了新的文献求助30
13秒前
小晓发布了新的文献求助10
13秒前
科研通AI2S应助Andema采纳,获得10
13秒前
15秒前
15秒前
18秒前
18秒前
20秒前
李健应助假发君采纳,获得10
21秒前
22秒前
KIORking发布了新的文献求助10
22秒前
落忆发布了新的文献求助10
22秒前
tengfei完成签到 ,获得积分10
22秒前
yinle发布了新的文献求助10
23秒前
23秒前
Ma发布了新的文献求助10
23秒前
23秒前
shensiang完成签到,获得积分10
24秒前
2116564发布了新的文献求助10
24秒前
25秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
‘Unruly’ Children: Historical Fieldnotes and Learning Morality in a Taiwan Village (New Departures in Anthropology) 400
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3989444
求助须知:如何正确求助?哪些是违规求助? 3531531
关于积分的说明 11254250
捐赠科研通 3270191
什么是DOI,文献DOI怎么找? 1804901
邀请新用户注册赠送积分活动 882105
科研通“疑难数据库(出版商)”最低求助积分说明 809174