title: Biophysical characterization of in vitro bound Streptomyces peucetius daunorubicin-serine protease complex.
abstract: A serine protease of Streptomyces peucetius is found in association with daunorubicin in the culture filtrate and co-purifies as a complex as reported earlier by us (Dubey et al., 2013). The same protease was purified without drug attachment from dpsA(-) mutant of S. peucetius, which does not produce daunorubicin. Drug-protein complex was made in vitro by mixing daunorubicin and the protease. Spectral analysis and circular dichroism (CD) analysis were employed to determine the interaction between daunorubicin and the protease. Our study showed that interaction of daunorubicin with the protease affects the spectral characteristics of the drug and changes the secondary structure of the protein. Thin layer chromatography (TLC) analysis showed that the drug-protein interaction results in partial conversion of the drug to aglyconic form. The complex formation implies sequestration of the drug when it attains potentially lethal level in the extracellular milieu of S. peucetius culture.
authors.affiliation: Department of Genetic Engineering, School of Biotechnology, Madurai Kamaraj University, Madurai 625021, India. Electronic address: bharadhwajrashmi@gmail.com.
authors.forename: Rashmi
authors.initials: R
authors.lastname: Dubey
authors.affiliation: Department of Genetic Engineering, School of Biotechnology, Madurai Kamaraj University, Madurai 625021, India. Electronic address: ranjan_mku@yahoo.com.
authors.forename: Ranjan
authors.initials: R
authors.lastname: Prasad
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000737:chemistry / Q000378:metabolism; D003630:Daunorubicin / Q000737:chemistry / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm; D006868:Hydrolysis; D009154:Mutation; D011485:Protein Binding; D057057:Serine Proteases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics
chemical_list: D000903:Antibiotics, Antineoplastic; D057057:Serine Proteases; D003630:Daunorubicin
keywords: Daunorubicin–protease complex; Self-resistance; Streptomyces peucetius