Identification of Cisplatin-Binding Proteins Using Agarose Conjugates of Platinum Compounds

Takatoshi Karasawa, Martha Sibrian-Vazquez, Robert M. Strongin, Peter Steyger

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Cisplatin is widely used as an antineoplastic drug, but its ototoxic and nephrotoxic side-effects, as well as the inherent or acquired resistance of some cancers to cisplatin, remain significant clinical problems. Cisplatin's selectivity in killing rapidly proliferating cancer cells is largely dependent on covalent binding to DNA via cisplatin's chloride sites that had been aquated. We hypothesized that cisplatin's toxicity in slowly proliferating or terminally differentiated cells is primarily due to drug-protein interactions, instead of drug-DNA binding. To identify proteins that bind to cisplatin, we synthesized two different platinum-agarose conjugates, one with two amino groups and another with two chlorides attached to platinum that are available for protein binding, and conducted pull-down assays using cochlear and kidney cells. Mass spectrometric analysis on protein bands after gel electrophoresis and Coomassie blue staining identified several proteins, including myosin IIA, glucose-regulated protein 94 (GRP94), heat shock protein 90 (HSP90), calreticulin, valosin containing protein (VCP), and ribosomal protein L5, as cisplatin-binding proteins. Future studies on the interaction of these proteins with cisplatin will elucidate whether these drug-protein interactions are involved in ototoxicity and nephrotoxicity, or contribute to tumor sensitivity or resistance to cisplatin treatment.

Original languageEnglish (US)
Article numbere66220
JournalPLoS One
Volume8
Issue number6
DOIs
StatePublished - Jun 3 2013

Fingerprint

Platinum Compounds
platinum
cisplatin
Sepharose
agarose
Cisplatin
binding proteins
Carrier Proteins
proteins
Proteins
Platinum
Drug Interactions
drugs
Chlorides
chlorides
Nonmuscle Myosin Type IIA
Pharmaceutical Preparations
calreticulin
Calreticulin
HSP90 Heat-Shock Proteins

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Identification of Cisplatin-Binding Proteins Using Agarose Conjugates of Platinum Compounds. / Karasawa, Takatoshi; Sibrian-Vazquez, Martha; Strongin, Robert M.; Steyger, Peter.

In: PLoS One, Vol. 8, No. 6, e66220, 03.06.2013.

Research output: Contribution to journalArticle

Karasawa, Takatoshi ; Sibrian-Vazquez, Martha ; Strongin, Robert M. ; Steyger, Peter. / Identification of Cisplatin-Binding Proteins Using Agarose Conjugates of Platinum Compounds. In: PLoS One. 2013 ; Vol. 8, No. 6.
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