Combination of two-dimensional gel electrophoresis and a fluorescent carboxyfluorescein-diacetate-labeled cisplatin analogue allows the identification of intracellular cisplatin-protein adducts

Sandra Kotz; Maximilian Kullmann; Barbara Crone; Ganna V Kalayda; Ulrich Jaehde; Sabine Metzger

doi:10.1002/elps.201500188

Combination of two-dimensional gel electrophoresis and a fluorescent carboxyfluorescein-diacetate-labeled cisplatin analogue allows the identification of intracellular cisplatin-protein adducts

Electrophoresis. 2015 Nov;36(21-22):2811-2819. doi: 10.1002/elps.201500188. Epub 2015 Sep 22.

Authors

Sandra Kotz¹, Maximilian Kullmann², Barbara Crone³, Ganna V Kalayda², Ulrich Jaehde², Sabine Metzger^{1

4}

Affiliations

¹ Biocenter MS Platform, Department of Biology, University of Cologne, Cologne, Germany.
² Department of Clinical Pharmacy, Institute of Pharmacy, University of Bonn, Bonn, Germany.
³ Institute of Inorganic and Analytical Chemistry, University of Münster, Münster, Germany.
⁴ IUF-Leibniz Research Institute for Environmental Medicine, IUF, Düsseldorf, Germany.

PMID: 26250553
DOI: 10.1002/elps.201500188

Abstract

Cisplatin is one of the most widely used anticancer agents, but a major problem for successful chemotherapy is the development of drug resistance of tumor cells against cisplatin. Resistance to cisplatin is a multifactorial problem. A method to detect and identify intracellular cisplatin-protein adducts was developed using a fluorescent carboxyfluorescein-diacetate-labeled cisplatin analogue (CFDA-cisplatin), 2DE, and ESI-MS/MS. We identified several CFDA-cisplatin-protein adducts including members of the protein disulfide isomerase family (PDI). These are the first results of the detection of intracellular CFDA-cisplatin-protein adducts, which may help to understand the resistance mechanism of cisplatin.

Keywords: Carboxyfluorescein-diacetate-labeled cisplatin analogue; Cisplatin; Cisplatin-protein adducts; Protein marker grid; Two-dimensional gel electrophoresis.