Unknown

Dataset Information

0

Kinetic and Structural Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag.


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.

SUBMITTER: Wilhelm J 

PROVIDER: S-EPMC8388125 | biostudies-literature |

REPOSITORIES: biostudies-literature

Similar Datasets

| S-EPMC3920298 | biostudies-other
| S-EPMC3808294 | biostudies-literature
| S-EPMC7596905 | biostudies-literature
| S-EPMC2850560 | biostudies-literature
| S-EPMC6474801 | biostudies-literature
| S-EPMC4800872 | biostudies-literature
| S-EPMC7434562 | biostudies-literature
| S-EPMC6044709 | biostudies-literature
| S-EPMC3199291 | biostudies-literature
| S-EPMC4638144 | biostudies-literature