Universität zu Köln

Kotz, Sandra, Kullmann, Maximilian, Kalayda, Ganna V., Dyballa-Rukes, Nadine, Jaehde, Ulrich and Metzger, Sabine (2018). Optimized two-dimensional gel electrophoresis in an alkaline pH range improves the identification of intracellular CFDA-cisplatin-protein adducts in ovarian cancer cells. Electrophoresis, 39 (12). S. 1488 - 1497. HOBOKEN: WILEY. ISSN 1522-2683

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Abstract

Intracellular binding of cisplatin to proteins has been associated with acquired resistance to chemotherapy. In our previous study we established an analytical method for the identification of intracellular cisplatin-binding proteins. The method used a fluorescent carboxyfluorescein-diacetate-labeled cisplatin analogue (CFDA-cisplatin), two-dimensional gel electrophoresis (2DE) and mass spectrometry, which allows detecting and identifying intracellular CFDA-cisplatin-containing protein adducts in the acidic pH range (pH 4-7). Based on this analytical method we extended the identification of intracellular cisplatin-protein adducts to the alkaline pH range (pH 6-10) giving chance to discover new important binding partners. 2DE analysis of alkaline proteins is challenging due to the difficult separation of basic proteins during the isoelectric focusing (IEF). The establishment of an optimized IEF protocol for basic proteins enabled us to identify several intracellular CFDA-cisplatin-binding proteins including enzymes of the glucose and serine metabolism like alpha enolase and D-3-phosphoglycerate 1-dehydrogenase.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Kotz, Sandra UNSPECIFIED UNSPECIFIED UNSPECIFIED Kullmann, Maximilian UNSPECIFIED UNSPECIFIED UNSPECIFIED Kalayda, Ganna V. UNSPECIFIED UNSPECIFIED UNSPECIFIED Dyballa-Rukes, Nadine UNSPECIFIED UNSPECIFIED UNSPECIFIED Jaehde, Ulrich UNSPECIFIED UNSPECIFIED UNSPECIFIED Metzger, Sabine UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-184542
DOI:	10.1002/elps.201700377
Journal or Publication Title:	Electrophoresis
Volume:	39
Number:	12
Page Range:	S. 1488 - 1497
Date:	2018
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1522-2683
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ELONGATION-FACTOR 1A; PHOSPHOGLYCERATE DEHYDROGENASE; PLATINUM COMPLEXES; BASIC-PROTEINS; ALPHA-ENOLASE; CURRENT STATE; UP-REGULATION; MECHANISMS; METABOLISM; RESISTANCE Multiple languages Biochemical Research Methods; Chemistry, Analytical Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/18454