Spectroscopic and Calorimetric Approach to Understand the Molecular Basis of Self-Association of Aureolic Acid Antibiotic, Chromomycin A3

DOI: 10.4236/ojbiphy.2014.42008   PDF   HTML     3,168 Downloads   5,149 Views  


Chromomycin A3 (CHR, pKa = 7.0), an aureolic acid group of antitumor antibiotic, undergoes self-association in aqueous solution in neutral and anionic forms. Self-association processes of neutral and anionic CHR have been studied in pH 5.0 and pH 9.0, respectively using different spectroscopic methods such as absorbance, fluorescence, CD, NMR and isothermal titration calorimetry (ITC). Results from these studies reveal that at low concentration (<10 μM), CHR exists predominantly in dimeric state for both neutral and anionic forms. With the increase of concentration, dimers further aggregate to form trimer and teramer in the following steps: (CHR)2 + CHR (CHR)3 and (CHR)3 + CHR (CHR)4. Analysis of NMR spectra of 100 μM and 1 mM CHR indicates that the self-association of CHR (neutral and anionic form) is most likely to happen via hydrophobic interaction involving the sugar moieties and surrounding water molecules. Calorimetric studies indicate that self-association of both anionic and neutral CHR is entropy driven. These observations imply that sugar substituents play a major role in their state of aggregation after biosynthesis from a gene cluster. The self-association features of the antibiotic have been compared with those of Mithramycin, an antibiotic of the same group.

Share and Cite:

Dutta, S. , Lahiri, S. and Dasgupta, D. (2014) Spectroscopic and Calorimetric Approach to Understand the Molecular Basis of Self-Association of Aureolic Acid Antibiotic, Chromomycin A3. Open Journal of Biophysics, 4, 66-82. doi: 10.4236/ojbiphy.2014.42008.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Calabresi, P., Chabner, B.A., Hardman, J.G. and Limbard, L.E. (1991) Goodman and Gilman’s “The Pharmacological Basis of Therapeutics: Chemotherapy of Neoplastic Diseases”. Macmillan, New York, 1225-1269.
[2] Lombo, F., Menendez, N., Salas, J.A. and Mendez, C. (2006) The Aureolic Acid Family of Antitumor Compounds: Structure, Mode of Action, Biosynthesis, and Novel Derivatives. Applied Microbiology and Biotechnology, 73, 1-14.
[3] Reynolds, R.D., Fisher, J.I., Jensen, P.A., Pajak, T.F. and Bateman, J.R. (1976) Phase I Alternate-Day Dose Study of Chromomycin A3. Cancer Treatment Reports, 60, 1251-1255.
[4] Sahar, E. and Latt, S.A. (1978) Enhancement of Banding Patterns in Human Metaphase Chromosomes by Energy Transfer. Proceedings of the National Academy of Sciences USA, 75, 5650-5654.
[5] Goldberg, I.H. and Friedman, P. (1971) Antibiotics and Nucleic Acids. Annual Review of Biochemistry, 40, 775-810.
[6] Aich, P., Sen, R. and Dasgupta, D. (1992) Role of Magnesium Ion in the Interaction between Chromomycin A3 and DNA: Binding of Chromomycin A3-Mg2+ Complexes with DNA. Biochemistry, 31, 2988-2997.
[7] Snyder, R.C., Ray, R., Blume, S. and Miller, D.M. (1991) Mithramycin Blocks Transcriptional Initiation of the c-myc P1 and P2 Promoters. Biochemistry, 30, 4290-4297.
[8] Fibach, E., Bianchi, N., Borgatti, M., Prus, E. and Gambari, R. (2003) Mithramycin Induces Fetal Hemoglobin Production in Normal and Thalassemic Human Erythroid Precursor Cells. Blood, 102, 1276-1281.
[9] Devi, P.G., Chakraborty, P.K. and Dasgupta, D. (2009) Inhibition of a Zn (II)-Containing Enzyme, Alcohol Dehydrogenase, by Anticancer Antibiotics, Mithramycin and Chromomycin A3. Journal of Biological Inorganic Chemistry, 14, 347- 359.
[10] Menendez, N., Nur-e-Alam, M., Brana, A.F., Rohr, J.A. and Mendez, C. (2004) Tailoring Modification of Deoxysugars during Biosynthesis of the Antitumour Drug Chromomycin A3 by Streptomyces griseus ssp. Griseus. Molecular Microbiology, 53, 903-915.
[11] Bosserman, M.A., Florez, A.B., Shaaban, K.A., Brana, A.F., Salas, J.A., Mendez, C. and Rohr, J. (2011) Characterization of the Terminal Activation Step Catalyzed by Oxygenase CmmOIV of the Chromomycin Biosynthetic Pathway from Streptomyces griseus. Biochemistry, 50, 1421-1428. http://dx.doi.org/10.1021/bi1016205
[12] Beam, M.P., Bosserman, M.A., Noinaj, N., Wehenkel, M. and Rohr, J. (2009) Crystal Structure of Baeyer-Villiger Monooxygenase MtmOIV, the Key Enzyme of the Mithramycin Biosynthetic Pathway. Biochemistry, 48, 4476-4487.
[13] Lahiri, S., Devi, P.G., Majumder, P., Das, S. and Dasgupta, D. (2008) Self-Association of the Anionic Form of the DNA-Binding Anticancer Drug Mithramycin. The Journal of Physical Chemistry B, 112, 3251-3258. http://dx.doi.org/10.1021/jp710503g
[14] Nayak, R., Sirsi, M. and Podder, S.K. (1975) Mode of Action of Antitumour Antibiotics: Spectrophotometric Studies on the Interaction of Chromomycin A3 with DNA and Chromatin of Normal and Neoplastic Tissue. Biochimica et Biophysica Acta (BBA)-Nucleic Acids and Protein Synthesis, 378, 195-204.
[15] Martin, S.R. (1980) Absorption and Circular Dichroic Spectral Studies on the Self-Association of Daunorubicin. Biopolymers, 19, 713-721. http://dx.doi.org/10.1002/bip.1980.360190318
[16] Gao, X. and Patel, D.J. (1989) Solution Structure of the Chromomycin-DNA Complex. Biochemistry, 28, 751-762. http://dx.doi.org/10.1021/bi00428a051
[17] Gao, X. and Patel, D.J. (1990) Chromomycin Dimer-DNA Oligomer Complexes. Sequence Selectivity and Divalent Cation Specificity. Biochemistry, 29, 10940-10956. http://dx.doi.org/10.1021/bi00501a012
[18] Devi, P.G., Pal, S., Banerjee, R. and Dasgupta, D. (2007) Association of Antitumor Antibiotics, Mithramycin and Chromomycin, with Zn(II). Journal of Inorganic Biochemistry, 101, 127-137. http://dx.doi.org/10.1016/j.jinorgbio.2006.08.018
[19] Lahiri, S., Takao, T., Devi, P.G., Ghosh, S., Ghosh, A., Dasgupta, A. and Dasgupta, D. (2012) Association of Aureolic Acid Antibiotic, Chromomycin A3 with Cu2+ and Its Negative Effect upon DNA Binding Property of the Antibiotic. Biometals, 25, 435-450.
[20] Chaires, J.B., Dattagupta, N. and Crothers, D.M. (1982) Self-Association of Daunomycin. Biochemistry, 21, 3927-3932. http://dx.doi.org/10.1021/bi00260a004
[21] Kikuchi, T., Ito, N., Suzuki, M., Kusai, A., Iseki, K. and Sasaki, H. (2005) Self-Association Properties of 4-[1- Hydroxy-1-Methylethyl]-2-Propyl-1-[4-[2-[Tetrazole-5-yl]Phenyl]Phenyl] Methylimidazole-5-Carboxylic Acid Mono- hydrate (CS-088), an Antiglaucoma Ophthalmic Agent. International Journal of Pharmaceutics, 299, 100-106. http://dx.doi.org/10.1016/j.ijpharm.2005.04.035
[22] Lewis, R.J., Hughes, R.A., Alcaraz, L., Thompson, S.P. and Moody, C.J. (2006) Solution Structures of Thiopeptide Antibiotics. Chemical Communications, 40, 4215-4217. http://dx.doi.org/10.1039/b609282a
[23] Grijalba, M.T., Cheron, M., Borowski, E., Bolard, J. and Schreier, S. (2006) Modulation of Polyene Antibiotics Self-Association by Ions from the Hofmeister Series. Biochimica et Biophysica Acta (BBA)—General Subjects, 1760, 973- 979. http://dx.doi.org/10.1016/j.bbagen.2006.02.004
[24] Veselkov, D., Lantushenko, A., Davies, D. and Veselkov, A. (2002) The Self-Association of Antibiotic Actinocyl-Bis (3-Dimethylaminopropylamine) in Aqueous Solution: A 1 H NMR Analysis. Russian Journal of Bioorganic Chemistry, 28, 342-347. http://dx.doi.org/10.1023/A:1019556211221
[25] Patel, T.R., Harding, S.E., Ebringerova, A., Deszczynski, M., Hromadkova, Z., Togola, A., Paulsen, B.S., Morris, G.A. and Rowe, A.J. (2007) Weak Self-Association in a Carbohydrate System. Biophys Journal, 93, 741-749. http://dx.doi.org/10.1529/biophysj.106.100891
[26] Santacroce, P.V. and Basu, A. (2004) Studies of the Carbohydrate-Carbohydrate Interaction between Lactose and GM3 Using Langmuir Monolayers and Glycolipid Micelles. Glycoconjugate Journal, 21, 89-95. http://dx.doi.org/10.1023/B:GLYC.0000044841.12706.12
[27] Yano, Y., Tanaka, K., Doi, Y. and Janado, M. (1988) The Polystyrene Affinity of Methylglycosides, Deoxysugars and Glucooligosaccharides. Journal of Solution Chemistry, 17, 347-358.
[28] Singh, B. And Gupta, R.S. (1985) Species-Specific Differences in the Toxicity and Mutagenicity of the Anticancer Drugs Mithramycin, Chromomycin A3, and Olivomycin. Cancer Research, 45, 2813-2820.
[29] Chan, J., Khan, S.N., Harvey, I., Merrick, W. and Pelletier, J. (2004) Eukaryotic Protein Synthesis Inhibitors Identified by Comparison of Cytotoxicity Profiles. RNA Society, 10, 528-543. http://dx.doi.org/10.1261/rna.5200204
[30] Chakrabarti, S., Bhattacharyya, D. and Dasgupta, D. (2001) Structural Basis of DNA Recognition by Anticancer Antibiotics, Chromomycin A3, and Mithramycin: Roles of Minor Groove Width and Ligand Flexibility. Biopolymers, 56, 85-95.
[31] Chakrabarti, S., Bhattacharyya, B. and Dasgupta, D. (2002) Interaction of Mithramycin and Chromomycin A3 with d (TAGCTAGCTA)2: Role of Sugars in Antibiotic-DNA Recognition. The Journal of Physical Chemistry B, 106, 6947-6953. http://dx.doi.org/10.1021/jp014710i

comments powered by Disqus

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.