hyt13

Amino Acid Sequence

3D Structure (/structures/AF-P0A3G2-F1-model_v6.cif)

Function

Catalyzes hydrolytic cleavage of carbon-halogen bonds in halogenated aliphatic compounds, leading to the formation of the corresponding primary alcohols, halide ions and protons. Expresses halogenase activity against 1-chloroalkanes of chain length C3 to C10, and also shows a very weak activity with 1,2-dichloroethane.

Mechanism

The ubiquitin-proteasome pathway (UPP) constitutes the principal pathway for protein degradation in eukaryotic cells, mediating the breakdown of target proteins through ubiquitin tagging and subsequent proteasomal degradation. In an ATP-dependent manner, the ubiquitin-activating enzyme (E1) activates ubiquitin (Ub) and conjugates it to a ubiquitin-conjugating enzyme (E2). E3 ligase subsequently catalyzes the transfer of Ub to the target protein, leading to the formation of a polyubiquitin chain that completes the ubiquitination mark. The 19S regulatory particle of the 26S proteasome recognizes this ubiquitination signal and, after deubiquitination, mediates the ATP-dependent delivery of the substrate into the 20S proteasome for degradation.

Description

Consistent with our hypothesis that hydrophobic tagging mimics a partially denatured protein state and that the protein is ultimately delivered to the proteasome for degradation, inclusion of proteasome inhibitors MG132 and YU101 blocked HyT13 mediated degradation (Fig. 2c).

Description

The IC50 of HyT13 was determined to be 21 nM (Supplementary Results, Supplementary Fig. 1). Supplementary Figure 1. The concentration curve of HyT13. Flp-In 293 cells expressing HA-EGFP-HaloTag were treated with indicated concentrations of HyT13 for 24 hours. The lysates were probed with anti-HA and anti-β actin antibodies, with β-actin serving as a loading control. Shown is quantification of three separate experiments, with error bars representing SEM.