6LZ2
Crystal structure of a thermostable green fluorescent protein (TGP) with a synthetic nanobody (Sb44)
Summary for 6LZ2
| Entry DOI | 10.2210/pdb6lz2/pdb |
| Descriptor | Thermostable green fluorescent protein, synthetic nanobody (sybody) 44 against the thermostable green fluorescent protein (TGP), SODIUM ION, ... (7 entities in total) |
| Functional Keywords | complex, gfp, nanobody, single-chain antibody, sybody, synthetic antibody, tgp, thermostable green fluorescent protein, fluorescent protein |
| Biological source | Galaxea fascicularis More |
| Total number of polymer chains | 4 |
| Total formula weight | 85122.70 |
| Authors | Cai, H.,Yao, H.,Li, T.,Hutter, C.,Tang, Y.,Li, Y.,Seeger, M.,Li, D. (deposition date: 2020-02-17, release date: 2020-12-23, Last modification date: 2024-11-13) |
| Primary citation | Cai, H.,Yao, H.,Li, T.,Hutter, C.A.J.,Li, Y.,Tang, Y.,Seeger, M.A.,Li, D. An improved fluorescent tag and its nanobodies for membrane protein expression, stability assay, and purification. Commun Biol, 3:753-753, 2020 Cited by PubMed Abstract: Green fluorescent proteins (GFPs) are widely used to monitor membrane protein expression, purification, and stability. An ideal reporter should be stable itself and provide high sensitivity and yield. Here, we demonstrate that a coral (Galaxea fascicularis) thermostable GFP (TGP) is by such reasons an improved tag compared to the conventional jellyfish GFPs. TGP faithfully reports membrane protein stability at temperatures near 90 °C (20-min heating). By contrast, the limit for the two popular GFPs is 64 °C and 74 °C. Replacing GFPs with TGP increases yield for all four test membrane proteins in four expression systems. To establish TGP as an affinity tag for membrane protein purification, several high-affinity synthetic nanobodies (sybodies), including a non-competing pair, are generated, and the crystal structure of one complex is solved. Given these advantages, we anticipate that TGP becomes a widely used tool for membrane protein structural studies. PubMed: 33303987DOI: 10.1038/s42003-020-01478-z PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.03 Å) |
Structure validation
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