Target protein deglycosylation in living cells by a nanobody-fused split O-GlcNAcase
O-linked N -acetylglucosamine ( O -GlcNAc) is an essential and dynamic post-translational modification that is presented on thousands of nucleocytoplasmic proteins. Interrogating the role of O -GlcNAc on a single target protein is crucial, yet challenging to perform in cells. Herein, we developed a...
Gespeichert in:
| Veröffentlicht in: | Nature chemical biology 2021-05, Vol.17 (5), p.593-600 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 600 |
|---|---|
| container_issue | 5 |
| container_start_page | 593 |
| container_title | Nature chemical biology |
| container_volume | 17 |
| creator | Ge, Yun Ramirez, Daniel H. Yang, Bo D’Souza, Alexandria K. Aonbangkhen, Chanat Wong, Stephanie Woo, Christina M. |
| description | O-linked
N
-acetylglucosamine (
O
-GlcNAc) is an essential and dynamic post-translational modification that is presented on thousands of nucleocytoplasmic proteins. Interrogating the role of
O
-GlcNAc on a single target protein is crucial, yet challenging to perform in cells. Herein, we developed a nanobody-fused split
O
-GlcNAcase (OGA) as an
O
-GlcNAc eraser for selective deglycosylation of a target protein in cells. After systematic cellular optimization, we identified a split OGA with reduced inherent deglycosidase activity that selectively removed
O
-GlcNAc from the desired target protein when directed by a nanobody. We demonstrate the generality of the nanobody-fused split OGA using four nanobodies against five target proteins and use the system to study the impact of
O
-GlcNAc on the transcription factors c-Jun and c-Fos. The nanobody-directed
O
-GlcNAc eraser provides a new strategy for the functional evaluation and engineering of
O
-GlcNAc via the selective removal of
O
-GlcNAc from individual proteins directly in cells.
Fusion of a split form of the protein
O
-GlcNAcase with nanobodies enables the targeted removal of
O
-GlcNAc protein modifications, providing a tool for probing the functional roles of specific
O
-GlcNAc modifications in a cellular context. |
| doi_str_mv | 10.1038/s41589-021-00757-y |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_8085020</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2518018871</sourcerecordid><originalsourceid>FETCH-LOGICAL-c474t-646a09c62acbbe0d174ba014066cdf8db9a7d890647b2d50201a2bebd2c1c16c3</originalsourceid><addsrcrecordid>eNp9kctuFDEQRS0EIiHwAyyQJTZsDGW3221vkKIIAlJENsPa8msaRx57sLsj9d_Tw4ThsWBVVtWpW766CL2k8JZCJ981TnupCDBKAIZ-IMsjdE77nhHOhXp8evdwhp61dgfQCUHlU3TWdUIKpug52mxMHcOE97VMIWbsw5gWV9qSzBRLxmsrxfuYR-xCSg3bBRucTS62-IVs5xY8bvsUJ3xLrpP7culMC8_Rk61JLbx4qBfo68cPm6tP5Ob2-vPV5Q1xfOATEVwYUE4w46wN4OnArQHKQQjnt9JbZQYvFQg-WOZ7YEANs8F65qijwnUX6P1Rdz_bXfAu5KmapPc17kxddDFR_z3J8Zsey72WIA9yq8CbB4Favs-hTXoX28GoyaHMTTOuVCel4gf09T_oXZlrXu1p1lMJVMqBrhQ7Uq6W1mrYnj5DQR9C08fQ9Bqa_hmaXtalV3_aOK38SmkFuiPQ1lEeQ_19-z-yPwBXQaQo</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2518018871</pqid></control><display><type>article</type><title>Target protein deglycosylation in living cells by a nanobody-fused split O-GlcNAcase</title><source>MEDLINE</source><source>Nature</source><source>SpringerNature Journals</source><creator>Ge, Yun ; Ramirez, Daniel H. ; Yang, Bo ; D’Souza, Alexandria K. ; Aonbangkhen, Chanat ; Wong, Stephanie ; Woo, Christina M.</creator><creatorcontrib>Ge, Yun ; Ramirez, Daniel H. ; Yang, Bo ; D’Souza, Alexandria K. ; Aonbangkhen, Chanat ; Wong, Stephanie ; Woo, Christina M.</creatorcontrib><description>O-linked
N
-acetylglucosamine (
O
-GlcNAc) is an essential and dynamic post-translational modification that is presented on thousands of nucleocytoplasmic proteins. Interrogating the role of
O
-GlcNAc on a single target protein is crucial, yet challenging to perform in cells. Herein, we developed a nanobody-fused split
O
-GlcNAcase (OGA) as an
O
-GlcNAc eraser for selective deglycosylation of a target protein in cells. After systematic cellular optimization, we identified a split OGA with reduced inherent deglycosidase activity that selectively removed
O
-GlcNAc from the desired target protein when directed by a nanobody. We demonstrate the generality of the nanobody-fused split OGA using four nanobodies against five target proteins and use the system to study the impact of
O
-GlcNAc on the transcription factors c-Jun and c-Fos. The nanobody-directed
O
-GlcNAc eraser provides a new strategy for the functional evaluation and engineering of
O
-GlcNAc via the selective removal of
O
-GlcNAc from individual proteins directly in cells.
Fusion of a split form of the protein
O
-GlcNAcase with nanobodies enables the targeted removal of
O
-GlcNAc protein modifications, providing a tool for probing the functional roles of specific
O
-GlcNAc modifications in a cellular context.</description><identifier>ISSN: 1552-4450</identifier><identifier>EISSN: 1552-4469</identifier><identifier>DOI: 10.1038/s41589-021-00757-y</identifier><identifier>PMID: 33686291</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>631/92/221 ; 631/92/458 ; 631/92/96 ; Antigens, Neoplasm - chemistry ; Antigens, Neoplasm - genetics ; Antigens, Neoplasm - metabolism ; Biochemical Engineering ; Biochemistry ; Biological Assay ; Bioorganic Chemistry ; c-Fos protein ; c-Jun protein ; Catalytic Domain ; Cell Biology ; Cell fusion ; Chemistry ; Chemistry and Materials Science ; Chemistry/Food Science ; Deglycosylation ; Drug Delivery Systems - methods ; Gene Expression ; Glycosylation ; HEK293 Cells ; Histone Acetyltransferases - chemistry ; Histone Acetyltransferases - genetics ; Histone Acetyltransferases - metabolism ; Humans ; Hyaluronoglucosaminidase - chemistry ; Hyaluronoglucosaminidase - genetics ; Hyaluronoglucosaminidase - metabolism ; Hydrolysis ; JNK Mitogen-Activated Protein Kinases - genetics ; JNK Mitogen-Activated Protein Kinases - metabolism ; Membrane Glycoproteins - genetics ; Membrane Glycoproteins - metabolism ; N-Acetylglucosamine ; Nanobodies ; Nuclear Pore Complex Proteins - genetics ; Nuclear Pore Complex Proteins - metabolism ; Optimization ; Plasmids - chemistry ; Plasmids - metabolism ; Post-translation ; Protein Binding ; Proteins ; Recombinant Fusion Proteins - chemistry ; Recombinant Fusion Proteins - genetics ; Recombinant Fusion Proteins - metabolism ; Single-Domain Antibodies - chemistry ; Single-Domain Antibodies - metabolism ; Sp1 Transcription Factor - genetics ; Sp1 Transcription Factor - metabolism ; Transcription factors ; Transcription Factors - genetics ; Transcription Factors - metabolism ; Transfection - methods</subject><ispartof>Nature chemical biology, 2021-05, Vol.17 (5), p.593-600</ispartof><rights>The Author(s), under exclusive licence to Springer Nature America, Inc. 2021</rights><rights>The Author(s), under exclusive licence to Springer Nature America, Inc. 2021.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-646a09c62acbbe0d174ba014066cdf8db9a7d890647b2d50201a2bebd2c1c16c3</citedby><cites>FETCH-LOGICAL-c474t-646a09c62acbbe0d174ba014066cdf8db9a7d890647b2d50201a2bebd2c1c16c3</cites><orcidid>0000-0001-8687-9105 ; 0000-0003-3534-7517 ; 0000-0003-2429-3218 ; 0000-0002-7378-1341 ; 0000-0003-3288-9951 ; 0000-0001-6769-5458 ; 0000-0001-9144-7841</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/s41589-021-00757-y$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/s41589-021-00757-y$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>230,314,780,784,885,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33686291$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ge, Yun</creatorcontrib><creatorcontrib>Ramirez, Daniel H.</creatorcontrib><creatorcontrib>Yang, Bo</creatorcontrib><creatorcontrib>D’Souza, Alexandria K.</creatorcontrib><creatorcontrib>Aonbangkhen, Chanat</creatorcontrib><creatorcontrib>Wong, Stephanie</creatorcontrib><creatorcontrib>Woo, Christina M.</creatorcontrib><title>Target protein deglycosylation in living cells by a nanobody-fused split O-GlcNAcase</title><title>Nature chemical biology</title><addtitle>Nat Chem Biol</addtitle><addtitle>Nat Chem Biol</addtitle><description>O-linked
N
-acetylglucosamine (
O
-GlcNAc) is an essential and dynamic post-translational modification that is presented on thousands of nucleocytoplasmic proteins. Interrogating the role of
O
-GlcNAc on a single target protein is crucial, yet challenging to perform in cells. Herein, we developed a nanobody-fused split
O
-GlcNAcase (OGA) as an
O
-GlcNAc eraser for selective deglycosylation of a target protein in cells. After systematic cellular optimization, we identified a split OGA with reduced inherent deglycosidase activity that selectively removed
O
-GlcNAc from the desired target protein when directed by a nanobody. We demonstrate the generality of the nanobody-fused split OGA using four nanobodies against five target proteins and use the system to study the impact of
O
-GlcNAc on the transcription factors c-Jun and c-Fos. The nanobody-directed
O
-GlcNAc eraser provides a new strategy for the functional evaluation and engineering of
O
-GlcNAc via the selective removal of
O
-GlcNAc from individual proteins directly in cells.
Fusion of a split form of the protein
O
-GlcNAcase with nanobodies enables the targeted removal of
O
-GlcNAc protein modifications, providing a tool for probing the functional roles of specific
O
-GlcNAc modifications in a cellular context.</description><subject>631/92/221</subject><subject>631/92/458</subject><subject>631/92/96</subject><subject>Antigens, Neoplasm - chemistry</subject><subject>Antigens, Neoplasm - genetics</subject><subject>Antigens, Neoplasm - metabolism</subject><subject>Biochemical Engineering</subject><subject>Biochemistry</subject><subject>Biological Assay</subject><subject>Bioorganic Chemistry</subject><subject>c-Fos protein</subject><subject>c-Jun protein</subject><subject>Catalytic Domain</subject><subject>Cell Biology</subject><subject>Cell fusion</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Chemistry/Food Science</subject><subject>Deglycosylation</subject><subject>Drug Delivery Systems - methods</subject><subject>Gene Expression</subject><subject>Glycosylation</subject><subject>HEK293 Cells</subject><subject>Histone Acetyltransferases - chemistry</subject><subject>Histone Acetyltransferases - genetics</subject><subject>Histone Acetyltransferases - metabolism</subject><subject>Humans</subject><subject>Hyaluronoglucosaminidase - chemistry</subject><subject>Hyaluronoglucosaminidase - genetics</subject><subject>Hyaluronoglucosaminidase - metabolism</subject><subject>Hydrolysis</subject><subject>JNK Mitogen-Activated Protein Kinases - genetics</subject><subject>JNK Mitogen-Activated Protein Kinases - metabolism</subject><subject>Membrane Glycoproteins - genetics</subject><subject>Membrane Glycoproteins - metabolism</subject><subject>N-Acetylglucosamine</subject><subject>Nanobodies</subject><subject>Nuclear Pore Complex Proteins - genetics</subject><subject>Nuclear Pore Complex Proteins - metabolism</subject><subject>Optimization</subject><subject>Plasmids - chemistry</subject><subject>Plasmids - metabolism</subject><subject>Post-translation</subject><subject>Protein Binding</subject><subject>Proteins</subject><subject>Recombinant Fusion Proteins - chemistry</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Single-Domain Antibodies - chemistry</subject><subject>Single-Domain Antibodies - metabolism</subject><subject>Sp1 Transcription Factor - genetics</subject><subject>Sp1 Transcription Factor - metabolism</subject><subject>Transcription factors</subject><subject>Transcription Factors - genetics</subject><subject>Transcription Factors - metabolism</subject><subject>Transfection - methods</subject><issn>1552-4450</issn><issn>1552-4469</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kctuFDEQRS0EIiHwAyyQJTZsDGW3221vkKIIAlJENsPa8msaRx57sLsj9d_Tw4ThsWBVVtWpW766CL2k8JZCJ981TnupCDBKAIZ-IMsjdE77nhHOhXp8evdwhp61dgfQCUHlU3TWdUIKpug52mxMHcOE97VMIWbsw5gWV9qSzBRLxmsrxfuYR-xCSg3bBRucTS62-IVs5xY8bvsUJ3xLrpP7culMC8_Rk61JLbx4qBfo68cPm6tP5Ob2-vPV5Q1xfOATEVwYUE4w46wN4OnArQHKQQjnt9JbZQYvFQg-WOZ7YEANs8F65qijwnUX6P1Rdz_bXfAu5KmapPc17kxddDFR_z3J8Zsey72WIA9yq8CbB4Favs-hTXoX28GoyaHMTTOuVCel4gf09T_oXZlrXu1p1lMJVMqBrhQ7Uq6W1mrYnj5DQR9C08fQ9Bqa_hmaXtalV3_aOK38SmkFuiPQ1lEeQ_19-z-yPwBXQaQo</recordid><startdate>20210501</startdate><enddate>20210501</enddate><creator>Ge, Yun</creator><creator>Ramirez, Daniel H.</creator><creator>Yang, Bo</creator><creator>D’Souza, Alexandria K.</creator><creator>Aonbangkhen, Chanat</creator><creator>Wong, Stephanie</creator><creator>Woo, Christina M.</creator><general>Nature Publishing Group US</general><general>Nature Publishing Group</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PCBAR</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-8687-9105</orcidid><orcidid>https://orcid.org/0000-0003-3534-7517</orcidid><orcidid>https://orcid.org/0000-0003-2429-3218</orcidid><orcidid>https://orcid.org/0000-0002-7378-1341</orcidid><orcidid>https://orcid.org/0000-0003-3288-9951</orcidid><orcidid>https://orcid.org/0000-0001-6769-5458</orcidid><orcidid>https://orcid.org/0000-0001-9144-7841</orcidid></search><sort><creationdate>20210501</creationdate><title>Target protein deglycosylation in living cells by a nanobody-fused split O-GlcNAcase</title><author>Ge, Yun ; Ramirez, Daniel H. ; Yang, Bo ; D’Souza, Alexandria K. ; Aonbangkhen, Chanat ; Wong, Stephanie ; Woo, Christina M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-646a09c62acbbe0d174ba014066cdf8db9a7d890647b2d50201a2bebd2c1c16c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>631/92/221</topic><topic>631/92/458</topic><topic>631/92/96</topic><topic>Antigens, Neoplasm - chemistry</topic><topic>Antigens, Neoplasm - genetics</topic><topic>Antigens, Neoplasm - metabolism</topic><topic>Biochemical Engineering</topic><topic>Biochemistry</topic><topic>Biological Assay</topic><topic>Bioorganic Chemistry</topic><topic>c-Fos protein</topic><topic>c-Jun protein</topic><topic>Catalytic Domain</topic><topic>Cell Biology</topic><topic>Cell fusion</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Chemistry/Food Science</topic><topic>Deglycosylation</topic><topic>Drug Delivery Systems - methods</topic><topic>Gene Expression</topic><topic>Glycosylation</topic><topic>HEK293 Cells</topic><topic>Histone Acetyltransferases - chemistry</topic><topic>Histone Acetyltransferases - genetics</topic><topic>Histone Acetyltransferases - metabolism</topic><topic>Humans</topic><topic>Hyaluronoglucosaminidase - chemistry</topic><topic>Hyaluronoglucosaminidase - genetics</topic><topic>Hyaluronoglucosaminidase - metabolism</topic><topic>Hydrolysis</topic><topic>JNK Mitogen-Activated Protein Kinases - genetics</topic><topic>JNK Mitogen-Activated Protein Kinases - metabolism</topic><topic>Membrane Glycoproteins - genetics</topic><topic>Membrane Glycoproteins - metabolism</topic><topic>N-Acetylglucosamine</topic><topic>Nanobodies</topic><topic>Nuclear Pore Complex Proteins - genetics</topic><topic>Nuclear Pore Complex Proteins - metabolism</topic><topic>Optimization</topic><topic>Plasmids - chemistry</topic><topic>Plasmids - metabolism</topic><topic>Post-translation</topic><topic>Protein Binding</topic><topic>Proteins</topic><topic>Recombinant Fusion Proteins - chemistry</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Single-Domain Antibodies - chemistry</topic><topic>Single-Domain Antibodies - metabolism</topic><topic>Sp1 Transcription Factor - genetics</topic><topic>Sp1 Transcription Factor - metabolism</topic><topic>Transcription factors</topic><topic>Transcription Factors - genetics</topic><topic>Transcription Factors - metabolism</topic><topic>Transfection - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ge, Yun</creatorcontrib><creatorcontrib>Ramirez, Daniel H.</creatorcontrib><creatorcontrib>Yang, Bo</creatorcontrib><creatorcontrib>D’Souza, Alexandria K.</creatorcontrib><creatorcontrib>Aonbangkhen, Chanat</creatorcontrib><creatorcontrib>Wong, Stephanie</creatorcontrib><creatorcontrib>Woo, Christina M.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Earth, Atmospheric & Aquatic Science Database</collection><collection>Materials Science Collection</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nature chemical biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ge, Yun</au><au>Ramirez, Daniel H.</au><au>Yang, Bo</au><au>D’Souza, Alexandria K.</au><au>Aonbangkhen, Chanat</au><au>Wong, Stephanie</au><au>Woo, Christina M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Target protein deglycosylation in living cells by a nanobody-fused split O-GlcNAcase</atitle><jtitle>Nature chemical biology</jtitle><stitle>Nat Chem Biol</stitle><addtitle>Nat Chem Biol</addtitle><date>2021-05-01</date><risdate>2021</risdate><volume>17</volume><issue>5</issue><spage>593</spage><epage>600</epage><pages>593-600</pages><issn>1552-4450</issn><eissn>1552-4469</eissn><abstract>O-linked
N
-acetylglucosamine (
O
-GlcNAc) is an essential and dynamic post-translational modification that is presented on thousands of nucleocytoplasmic proteins. Interrogating the role of
O
-GlcNAc on a single target protein is crucial, yet challenging to perform in cells. Herein, we developed a nanobody-fused split
O
-GlcNAcase (OGA) as an
O
-GlcNAc eraser for selective deglycosylation of a target protein in cells. After systematic cellular optimization, we identified a split OGA with reduced inherent deglycosidase activity that selectively removed
O
-GlcNAc from the desired target protein when directed by a nanobody. We demonstrate the generality of the nanobody-fused split OGA using four nanobodies against five target proteins and use the system to study the impact of
O
-GlcNAc on the transcription factors c-Jun and c-Fos. The nanobody-directed
O
-GlcNAc eraser provides a new strategy for the functional evaluation and engineering of
O
-GlcNAc via the selective removal of
O
-GlcNAc from individual proteins directly in cells.
Fusion of a split form of the protein
O
-GlcNAcase with nanobodies enables the targeted removal of
O
-GlcNAc protein modifications, providing a tool for probing the functional roles of specific
O
-GlcNAc modifications in a cellular context.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>33686291</pmid><doi>10.1038/s41589-021-00757-y</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0001-8687-9105</orcidid><orcidid>https://orcid.org/0000-0003-3534-7517</orcidid><orcidid>https://orcid.org/0000-0003-2429-3218</orcidid><orcidid>https://orcid.org/0000-0002-7378-1341</orcidid><orcidid>https://orcid.org/0000-0003-3288-9951</orcidid><orcidid>https://orcid.org/0000-0001-6769-5458</orcidid><orcidid>https://orcid.org/0000-0001-9144-7841</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1552-4450 |
| ispartof | Nature chemical biology, 2021-05, Vol.17 (5), p.593-600 |
| issn | 1552-4450 1552-4469 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_8085020 |
| source | MEDLINE; Nature; SpringerNature Journals |
| subjects | 631/92/221 631/92/458 631/92/96 Antigens, Neoplasm - chemistry Antigens, Neoplasm - genetics Antigens, Neoplasm - metabolism Biochemical Engineering Biochemistry Biological Assay Bioorganic Chemistry c-Fos protein c-Jun protein Catalytic Domain Cell Biology Cell fusion Chemistry Chemistry and Materials Science Chemistry/Food Science Deglycosylation Drug Delivery Systems - methods Gene Expression Glycosylation HEK293 Cells Histone Acetyltransferases - chemistry Histone Acetyltransferases - genetics Histone Acetyltransferases - metabolism Humans Hyaluronoglucosaminidase - chemistry Hyaluronoglucosaminidase - genetics Hyaluronoglucosaminidase - metabolism Hydrolysis JNK Mitogen-Activated Protein Kinases - genetics JNK Mitogen-Activated Protein Kinases - metabolism Membrane Glycoproteins - genetics Membrane Glycoproteins - metabolism N-Acetylglucosamine Nanobodies Nuclear Pore Complex Proteins - genetics Nuclear Pore Complex Proteins - metabolism Optimization Plasmids - chemistry Plasmids - metabolism Post-translation Protein Binding Proteins Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Single-Domain Antibodies - chemistry Single-Domain Antibodies - metabolism Sp1 Transcription Factor - genetics Sp1 Transcription Factor - metabolism Transcription factors Transcription Factors - genetics Transcription Factors - metabolism Transfection - methods |
| title | Target protein deglycosylation in living cells by a nanobody-fused split O-GlcNAcase |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-02T22%3A37%3A33IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Target%20protein%20deglycosylation%20in%20living%20cells%20by%20a%20nanobody-fused%20split%20O-GlcNAcase&rft.jtitle=Nature%20chemical%20biology&rft.au=Ge,%20Yun&rft.date=2021-05-01&rft.volume=17&rft.issue=5&rft.spage=593&rft.epage=600&rft.pages=593-600&rft.issn=1552-4450&rft.eissn=1552-4469&rft_id=info:doi/10.1038/s41589-021-00757-y&rft_dat=%3Cproquest_pubme%3E2518018871%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2518018871&rft_id=info:pmid/33686291&rfr_iscdi=true |