Development of Ubiquitin-Based Probe for Metalloprotease Deubiquitinases
| Autor: | Aysegul Sapmaz, Huib Ovaa, Lindsey Burggraaff, Gerard J. P. van Westen, Alessia Amore, Dharjath S. Hameed, Cornelis J Slingerland |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular Lysis Protein Conformation Metalloenzymes 010402 general chemistry 01 natural sciences Catalysis HeLa Ubiquitin Moiety Humans Chelating Agents chemistry.chemical_classification Metalloproteinase biology Deubiquitinating Enzymes 010405 organic chemistry Chemistry Biological activity General Chemistry biology.organism_classification 0104 chemical sciences Cell biology Zinc Enzyme biology.protein Chelates Fluorescent tag HeLa Cells |
| Zdroj: | Angewandte Chemie International Edition, 58(41), 14477-14482 Angewandte Chemie International Edition |
| Popis: | Deubiquitinases (DUBs) are a family of enzymes that regulate the ubiquitin signaling cascade by removing ubiquitin from specific proteins in response to distinct signals. DUBs that belong to the metalloprotease family (metalloDUBs) contain Zn2+ in their active sites and are an integral part of distinct cellular protein complexes. Little is known about these enzymes because of the lack of specific probes. Described here is a Ub‐based probe that contains a ubiquitin moiety modified at its C‐terminus with a Zn2+ chelating group based on 8‐mercaptoquinoline, and a modification at the N‐terminus with either a fluorescent tag or a pull‐down tag. The probe is validated using Rpn11, a metalloDUB found in the 26S proteasome complex. This probe binds to metalloDUBs and efficiently pulled down overexpressed metalloDUBs from a HeLa cell lysate. Such probes may be used to study the mechanism of metalloDUBs in detail and allow better understanding of their biochemical processes. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|