CD-spectroscopy as a powerful tool for investigating the mode of action of unmodified drugs in live cells.
Circular dichroism (CD) spectroscopy is a well-known method for the analysis of chiral chemical compounds and is often used for studying the structure and interaction of proteins, DNA and bioactive compounds in solution. Here we demonstrate that CD spectroscopy is also a powerful tool for investigating the cellular uptake and mode of action of drugs in live cells. By means of CD spectroscopy, we identified DNA as the cellular target of several novel anticancer agents based on the highly cytotoxic natural antibiotic CC-1065. Furthermore, time-dependent changes in the CD spectra of drug-treated cells enabled us to rationalize differences in drug cytotoxicity. The anticancer agents rapidly penetrate the cell membrane and bind to cellular DNA as their intracellular target. Thereby, the formation of a reversible noncovalent complex with the DNA is followed by a covalent binding of the drugs to the DNA and the more toxic compounds show a higher stability and a lower alkylation rate. Since no drug manipulation is necessary for this kind of investigation and achiral compounds bound to chiral biomolecules may also show induced CD signals, CD spectroscopy of live cells is not limited to the study of analogues of CC-1065. Thus, it constitutes a general approach for studying the mode of action of bioactive compounds on the cellular and molecular level.
Tietze
Lutz F
LF
Department of Organic and Biomolecular Chemistry, Georg-August-University of Göttingen, Tammannstrasse 2, 37077 Göttingen, Germany. ltietze@gwdg.de
Krewer
Birgit
B
Major
Felix
F
Schuberth
Ingrid
I
D000903:Antibiotics, Antineoplastic / Q000032:analysis / Q000493:pharmacokinetics / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D000080890:Duocarmycins; D006801:Humans; D007211:Indoles / Q000032:analysis / Q000493:pharmacokinetics / Q000494:pharmacology*; D015394:Molecular Structure
D000903:Antibiotics, Antineoplastic; D000080890:Duocarmycins; D007211:Indoles; C018782:CC 1065; D004247:DNA