Two-step bioorthogonal activity-based proteasome profiling using copper-free click reagents: a comparative study

Wouter A van der Linden; Nan Li; Sascha Hoogendoorn; Mark Ruben; Martijn Verdoes; Jun Guo; Geert-Jan Boons; Gijsbert A van der Marel; Bogdan I Florea; Herman S Overkleeft

doi:10.1016/j.bmc.2011.06.037

Two-step bioorthogonal activity-based proteasome profiling using copper-free click reagents: a comparative study

Bioorg Med Chem. 2012 Jan 15;20(2):662-6. doi: 10.1016/j.bmc.2011.06.037. Epub 2011 Jun 23.

Authors

Wouter A van der Linden¹, Nan Li, Sascha Hoogendoorn, Mark Ruben, Martijn Verdoes, Jun Guo, Geert-Jan Boons, Gijsbert A van der Marel, Bogdan I Florea, Herman S Overkleeft

Affiliation

¹ Leiden Institute of Chemistry and Netherlands Proteomics Centre, PO Box 9502, 2300 RA, Leiden, The Netherlands.

PMID: 21757357
DOI: 10.1016/j.bmc.2011.06.037

Abstract

The development and application of bioorthogonal two-step labeling techniques receives much attention. Employing bifunctional proteasome probe 2 the efficiency of two-step labeling of recently published biotinylated cyclooctynes 3-5 is compared to Staudinger-Bertozzi ligation in cell extracts and living cells. While cyclooctynes 3-5 react faster and at a much lower concentration then the Staudinger-Bertozzi benchmark, background labeling is considerable with these reagents.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Biotin / analogs & derivatives*
Biotin / chemistry
Cell Line
Click Chemistry*
Copper / chemistry
Cyclooctanes / chemistry*
Fluorescent Dyes / chemistry
Humans
Proteasome Endopeptidase Complex / chemistry
Proteasome Endopeptidase Complex / metabolism*

Substances

Cyclooctanes
Fluorescent Dyes
Biotin
Copper
Proteasome Endopeptidase Complex