Structure-based design of covalent nanobody binders for a thermostable green fluorescence protein
文献类型：期刊论文
| 作者 | Yue, Zhihao1,2; Li, Yanfang1; Cai, Hongmin1,3; Yao, Hebang1,4; Li, Dianfan1; Ni, Aimin1; Li, Tingting1 |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2025-08-01 |
| 卷号 | 57期号:8页码:1363-1370 |
| 关键词 | disulfide engineering rational design synthetic nanobody thermostable green fluorescence protein |
| ISSN号 | 1672-9145 |
| DOI | 10.3724/abbs.2024233 |
| 英文摘要 | The use of green fluorescence protein (GFP) has advanced numerous areas of life sciences. An ultra-thermostable GFP (TGP), engineered from a coral GFP, offers potential advantages over traditional jellyfish-derived GFP because of its high stability. However, owing to its later discovery, TGP lacks the extensive toolsets available for GFP, such as heavy chain-only antibody binders known as nanobodies. In this study, we report the crystal structure of TGP in complex with Sb92, a synthetic nanobody identified from a previous in vitro screening, revealing Sb92's precise three-dimensional epitope. This structural insight, alongside the previously characterized Sb44-TGP complex, allows us to rationally design disulfide bonds between the antigen and the antibody for tighter interactions. Using biochemical analysis, we identify two bridged complexes (TGP A18C-Sb44 V100C and TGP E118C-Sb92 S57C), with the TGP-Sb92 disulfide pair showing high resistance to reducing agents. Our study expands the toolkit available for TGP and should encourage its wider applications. |
| WOS关键词 | LIVING CELLS ; GFP ; MICROSCOPY ; REPORTER ; LOCALIZATION |
| 资助项目 | National Natural Science Foundation of China[32201000] ; National Natural Science Foundation of China[82151215] ; National Natural Science Foundation of China[32471246] ; Science and Technology Commission of Shanghai Municipality[22ZR1468300] ; Shanghai Postdoctoral Excellence Program[2021378] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biophysics |
| 语种 | 英语 |
| WOS记录号 | WOS:001609713300001 |
| 出版者 | SCIENCE PRESS |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/319814]  |
| 专题 | 国家级研究中心_原创新药研究全国重点实验室 |
| 通讯作者 | Ni, Aimin; Li, Tingting |
| 作者单位 | 1.Chinese Acad Sci, Ctr Excellence Mol Cell Sci, Shanghai Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai 201210, Peoples R China 4.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China |
| 推荐引用方式 GB/T 7714 | Yue, Zhihao,Li, Yanfang,Cai, Hongmin,et al. Structure-based design of covalent nanobody binders for a thermostable green fluorescence protein[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2025,57(8):1363-1370. |
| APA | Yue, Zhihao.,Li, Yanfang.,Cai, Hongmin.,Yao, Hebang.,Li, Dianfan.,...&Li, Tingting.(2025).Structure-based design of covalent nanobody binders for a thermostable green fluorescence protein.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,57(8),1363-1370. |
| MLA | Yue, Zhihao,et al."Structure-based design of covalent nanobody binders for a thermostable green fluorescence protein".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 57.8(2025):1363-1370. |
入库方式： OAI收割
来源：上海药物研究所
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