Global identification of O-GlcNAc transferase (OGT) interactors by a human proteome microarray and the construction of an OGT interactome
Rui-Ping Deng
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, P. R. China
State Key Laboratory of Oncogenes and Related Genes, Shanghai, P. R. China
These two authors contributed equally to this study.
Search for more papers by this authorXiang He
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, P. R. China
State Key Laboratory of Oncogenes and Related Genes, Shanghai, P. R. China
These two authors contributed equally to this study.
Search for more papers by this authorShu-Juan Guo
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, P. R. China
State Key Laboratory of Oncogenes and Related Genes, Shanghai, P. R. China
Search for more papers by this authorWei-Feng Liu
CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Chinese Academy of Sciences, Beijing, P. R. China
Search for more papers by this authorYong Tao
CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Chinese Academy of Sciences, Beijing, P. R. China
Search for more papers by this authorCorresponding Author
Sheng-Ce Tao
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, P. R. China
State Key Laboratory of Oncogenes and Related Genes, Shanghai, P. R. China
School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, P. R. China
Correspondence: Professor Sheng-Ce Tao, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, P. R. China
E-mail:[email protected]
Search for more papers by this authorRui-Ping Deng
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, P. R. China
State Key Laboratory of Oncogenes and Related Genes, Shanghai, P. R. China
These two authors contributed equally to this study.
Search for more papers by this authorXiang He
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, P. R. China
State Key Laboratory of Oncogenes and Related Genes, Shanghai, P. R. China
These two authors contributed equally to this study.
Search for more papers by this authorShu-Juan Guo
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, P. R. China
State Key Laboratory of Oncogenes and Related Genes, Shanghai, P. R. China
Search for more papers by this authorWei-Feng Liu
CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Chinese Academy of Sciences, Beijing, P. R. China
Search for more papers by this authorYong Tao
CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, Chinese Academy of Sciences, Beijing, P. R. China
Search for more papers by this authorCorresponding Author
Sheng-Ce Tao
Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, P. R. China
State Key Laboratory of Oncogenes and Related Genes, Shanghai, P. R. China
School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, P. R. China
Correspondence: Professor Sheng-Ce Tao, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, P. R. China
E-mail:[email protected]
Search for more papers by this authorColour Online: See the article online to view Figs. 2, 3 and 5 in colour.
Abstract
O-Linked β-N-acetylglucosamine (O-GlcNAcylation) is an important protein PTM, which is very abundant in mammalian cells. O-GlcNAcylation is catalyzed by O-GlcNAc transferase (OGT), whose substrate specificity is believed to be regulated through interactions with other proteins. There are a handful of known human OGT interactors, which is far from enough for fully elucidating the substrate specificity of OGT. To address this challenge, we used a human proteome microarray containing ∼17 000 affinity-purified human proteins to globally identify OGT interactors and identified 25 OGT-binding proteins. Bioinformatics analysis showed that these interacting proteins play a variety of roles in a wide range of cellular functions and are highly enriched in intra-Golgi vesicle-mediated transport and vitamin biosynthetic processes. Combining newly identified OGT interactors with the interactors identified prior to this study, we have constructed the first OGT interactome. Bioinformatics analysis suggests that the OGT interactome plays important roles in protein transportation/localization and transcriptional regulation. The novel OGT interactors that we identified in this study could serve as a starting point for further functional analysis. Because of its high-throughput and parallel analysis capability, we strongly believe that protein microarrays could be easily applied for the global identification of regulators for other key enzymes.
Supporting Information
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| Filename | Description |
|---|---|
| pmic7695-sup-0001-SuppMat.zip1.8 MB | Figure S1. Validation of the O-GlcNAcylation of PSAT1 and HAAO by immuno- precipitation and western blotting using an anti-O-GlcNAc antibody. (A). PSAT1, HAAO and the positive control OIP106 were immuno-precipitated and treated with PNGase F to remove the possible N-glycans. The proteins were then subjected for western blotting using an anti-O-GlcNAc antibody. (B). The cells were treated with PUGNAc, an O-GlcNAcase inhibitor before lysis and immuno-precipitation. Table S1. The list of the OGT interactors based on literatures. Table S2. Protein classification analysis of the OGT interactome. Table S3. Gene ontology analysis of the OGT interactome. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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