6Y7A
X-ray structure of the Haloalkane dehalogenase HaloTag7 labeled with a chloroalkane-tetramethylrhodamine fluorophore substrate
Summary for 6Y7A
| Entry DOI | 10.2210/pdb6y7a/pdb |
| Related | 6Y7B |
| Descriptor | Haloalkane dehalogenase, [9-[2-carboxy-5-[2-[2-(6-chloranylhexoxy)ethoxy]ethylcarbamoyl]phenyl]-6-(dimethylamino)xanthen-3-ylidene]-dimethyl-azanium, CHLORIDE ION, ... (5 entities in total) |
| Functional Keywords | haloalkane dehalogenase, halo, tag, halotag7, self-labeling protein, fluorophore, tetramethylrhodamine, hydrolase |
| Biological source | Rhodococcus sp. |
| Total number of polymer chains | 1 |
| Total formula weight | 33990.71 |
| Authors | Tarnawski, M.,Johnsson, K.,Hiblot, J. (deposition date: 2020-02-28, release date: 2021-03-31, Last modification date: 2024-10-16) |
| 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 (1.4 Å) |
Structure validation
Download full validation report
