6Y8P
Crystal structure of SNAP-tag labeled with a benzyl-tetramethylrhodamine fluorophore
Summary for 6Y8P
| Entry DOI | 10.2210/pdb6y8p/pdb |
| Descriptor | O6-alkylguanine-DNA alkyltransferase mutant, ZINC ION, [9-[2-carboxy-5-[(4-methylphenyl)methylcarbamoyl]phenyl]-6-(dimethylamino)xanthen-3-ylidene]-dimethyl-azanium, ... (5 entities in total) |
| Functional Keywords | snap-tag, self-labeling protein, tetramethylrhodamine, synthetic fluorophore, transferase |
| Biological source | Homo sapiens (Human) |
| Total number of polymer chains | 1 |
| Total formula weight | 19613.85 |
| Authors | Gotthard, G.,Tanzer, T.,Johnsson, K.,Hiblot, J. (deposition date: 2020-03-05, release date: 2021-03-31, Last modification date: 2024-10-23) |
| Primary citation | Wilhelm, J.,Kuhn, S.,Tarnawski, M.,Gotthard, G.,Tunnermann, J.,Tanzer, T.,Karpenko, J.,Mertes, N.,Xue, L.,Uhrig, U.,Reinstein, J.,Hiblot, J.,Johnsson, K. Kinetic and Structural Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag. Biochemistry, 60:2560-2575, 2021 Cited by PubMed Abstract: The self-labeling protein tags (SLPs) HaloTag7, SNAP-tag, and CLIP-tag allow the covalent labeling of fusion proteins with synthetic molecules for applications in bioimaging and biotechnology. To guide the selection of an SLP-substrate pair and provide guidelines for the design of substrates, we report a systematic and comparative study of the labeling kinetics and substrate specificities of HaloTag7, SNAP-tag, and CLIP-tag. HaloTag7 reaches almost diffusion-limited labeling rate constants with certain rhodamine substrates, which are more than 2 orders of magnitude higher than those of SNAP-tag for the corresponding substrates. SNAP-tag labeling rate constants, however, are less affected by the structure of the label than those of HaloTag7, which vary over 6 orders of magnitude for commonly employed substrates. Determining the crystal structures of HaloTag7 and SNAP-tag labeled with fluorescent substrates allowed us to rationalize their substrate preferences. We also demonstrate how these insights can be exploited to design substrates with improved labeling kinetics. PubMed: 34339177DOI: 10.1021/acs.biochem.1c00258 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.3 Å) |
Structure validation
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