Activity-Based Protein Profiling with Natural Product-Derived Chemical Probes in Human Cell Lysates

Susanne Zweerink; Tanja Pollmann; Sabrina Ninck; Farnusch Kaschani; Markus Kaiser

doi:10.1007/978-1-4939-6439-0_3

Activity-Based Protein Profiling with Natural Product-Derived Chemical Probes in Human Cell Lysates

Methods Mol Biol. 2017:1491:23-46. doi: 10.1007/978-1-4939-6439-0_3.

Authors

Susanne Zweerink¹, Tanja Pollmann¹, Sabrina Ninck¹, Farnusch Kaschani¹, Markus Kaiser²

Affiliations

¹ ZMB, Faculty of Biology, University of Duisburg-Essen, Universitätsstr. 2, 45117, Essen, Germany.
² ZMB, Faculty of Biology, University of Duisburg-Essen, Universitätsstr. 2, 45117, Essen, Germany. markus.kaiser@uni-due.de.

PMID: 27778279
DOI: 10.1007/978-1-4939-6439-0_3

Abstract

Bioreactive natural products represent versatile starting points for the development of structurally unique activity-based probes. In the present protocol, we describe the workflow for an activity-based protein profiling (ABPP) experiment with an alkyne-tagged natural product derivative. Our protocol includes experimental procedures for in vivo labeling, sample preparation and 2-step (click chemistry) visualization and sample preparation for mass spectrometry-based target identification.

Keywords: Activity-based protein profiling; Bioreactive natural products; Click probes; Target identification.

Publication types

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

MeSH terms

Biological Products / chemistry*
Click Chemistry
Hep G2 Cells
Humans
Molecular Probes / chemistry*
Proteins / chemistry*
Proteome

Substances

Biological Products
Molecular Probes
Proteins
Proteome