OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in <i>E. coli</i>
O-GlcNAcylation is a single glycosylation of GlcNAc mediated by OGT, which regulates the function of substrate proteins and is closely related to many diseases. However, a large number of O-GlcNAc-modified target proteins are costly, inefficient, and complicated to prepare. In this study, an OGT bin...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/5/2129 |
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| author | Yang Li Zelan Yang Jia Chen Yihao Chen Chengji Jiang Tao Zhong Yanting Su Yi Liang Hui Sun |
| author_facet | Yang Li Zelan Yang Jia Chen Yihao Chen Chengji Jiang Tao Zhong Yanting Su Yi Liang Hui Sun |
| author_sort | Yang Li |
| collection | DOAJ |
| description | O-GlcNAcylation is a single glycosylation of GlcNAc mediated by OGT, which regulates the function of substrate proteins and is closely related to many diseases. However, a large number of O-GlcNAc-modified target proteins are costly, inefficient, and complicated to prepare. In this study, an OGT binding peptide (OBP)-tagged strategy for improving the proportion of O-GlcNAc modification was established successfully in <i>E. coli</i>. OBP (P1, P2, or P3) was fused with target protein Tau as tagged Tau. Tau or tagged Tau was co-constructed with OGT into a vector expressed in <i>E. coli</i>. Compared with Tau, the O-GlcNAc level of P1Tau and TauP1 increased 4~6-fold. Moreover, the P1Tau and TauP1 increased the O-GlcNAc-modified homogeneity. The high O-GlcNAcylation on P1Tau resulted in a significantly slower aggregation rate than Tau in vitro. This strategy was also used successfully to increase the O-GlcNAc level of c-Myc and H2B. These results indicated that the OBP-tagged strategy was a successful approach to improve the O-GlcNAcylation of a target protein for further functional research. |
| first_indexed | 2024-03-11T07:18:05Z |
| format | Article |
| id | doaj.art-ebd44dfdd0134925936340897e70ad47 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-11T07:18:05Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-ebd44dfdd0134925936340897e70ad472023-11-17T08:12:23ZengMDPI AGMolecules1420-30492023-02-01285212910.3390/molecules28052129OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in <i>E. coli</i>Yang Li0Zelan Yang1Jia Chen2Yihao Chen3Chengji Jiang4Tao Zhong5Yanting Su6Yi Liang7Hui Sun8College of Life Sciences, Wuhan University, Wuhan 430072, ChinaCollege of Life Sciences, Wuhan University, Wuhan 430072, ChinaCollege of Life Sciences, Wuhan University, Wuhan 430072, ChinaCollege of Life Sciences, Wuhan University, Wuhan 430072, ChinaCollege of Life Sciences, Wuhan University, Wuhan 430072, ChinaCollege of Life Sciences, Wuhan University, Wuhan 430072, ChinaSchool of Basic Medical Sciences, Xianning Medical College, Hubei University of Science and Technology, Xianning 437100, ChinaCollege of Life Sciences, Wuhan University, Wuhan 430072, ChinaCollege of Life Sciences, Wuhan University, Wuhan 430072, ChinaO-GlcNAcylation is a single glycosylation of GlcNAc mediated by OGT, which regulates the function of substrate proteins and is closely related to many diseases. However, a large number of O-GlcNAc-modified target proteins are costly, inefficient, and complicated to prepare. In this study, an OGT binding peptide (OBP)-tagged strategy for improving the proportion of O-GlcNAc modification was established successfully in <i>E. coli</i>. OBP (P1, P2, or P3) was fused with target protein Tau as tagged Tau. Tau or tagged Tau was co-constructed with OGT into a vector expressed in <i>E. coli</i>. Compared with Tau, the O-GlcNAc level of P1Tau and TauP1 increased 4~6-fold. Moreover, the P1Tau and TauP1 increased the O-GlcNAc-modified homogeneity. The high O-GlcNAcylation on P1Tau resulted in a significantly slower aggregation rate than Tau in vitro. This strategy was also used successfully to increase the O-GlcNAc level of c-Myc and H2B. These results indicated that the OBP-tagged strategy was a successful approach to improve the O-GlcNAcylation of a target protein for further functional research.https://www.mdpi.com/1420-3049/28/5/2129OGTO-GlcNAcTauOBP-tagged strategy |
| spellingShingle | Yang Li Zelan Yang Jia Chen Yihao Chen Chengji Jiang Tao Zhong Yanting Su Yi Liang Hui Sun OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in <i>E. coli</i> Molecules OGT O-GlcNAc Tau OBP-tagged strategy |
| title | OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in <i>E. coli</i> |
| title_full | OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in <i>E. coli</i> |
| title_fullStr | OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in <i>E. coli</i> |
| title_full_unstemmed | OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in <i>E. coli</i> |
| title_short | OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in <i>E. coli</i> |
| title_sort | ogt binding peptide tagged strategy increases protein o glcnacylation level in i e coli i |
| topic | OGT O-GlcNAc Tau OBP-tagged strategy |
| url | https://www.mdpi.com/1420-3049/28/5/2129 |
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