title: Uptake and metabolism of fluorescent steroids by mycobacterial cells.
abstract: Fluorescent steroids BODIPY-cholesterol (BPCh) and 7-nitrobenzoxadiazole-4-amino-(NBD)-labeled 22-NBD-chelesterol (22NC) as well as synthesized 20-(NBD)-pregn-5-en-3β-ol (20NP) were found to undergo bioconversions by Mycobacterium tuberculosis H37Rv and M. smegmatis mc2155. The major fluorescent products were determined to be 4-en-3-one derivatives of the compounds. Degradation of NBD fluorophore was also detected in the cases of 22NC and 20NP, but neither NBD degradation nor steroidal part modification were observed for the synthesized 3-(NBD)-cholestane. Mycobacterial 3β-hydroxysteroid dehydrogenases were concluded to be responsible for the formation of the 4-en-3-one derivatives. All the compounds tested were found to cause staining both membrane lipids and cytosolic lipid droplets when incubated with mycobacteria in different manner, demonstrating ability of the steroids to reside in the compartments. The findings reveal a potential of the compounds for monitoring of steroid interactions with mycobacteria and provide information for design of new probes for this purpose.
authors.affiliation: Research Institute for Physical Chemical Problems, Belarusian State University, Minsk, Belarus; Faculty of Chemistry, Belarusian State University, Minsk, Belarus. Electronic address: yaroslav82@tut.by.
authors.forename: Yaroslav
authors.initials: Y
authors.lastname: Faletrov
authors.affiliation: Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland. Electronic address: abrzostek@cbm.pan.pl.
authors.forename: Anna
authors.initials: A
authors.lastname: Brzostek
authors.affiliation: Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland. Electronic address: rplocinska@cbm.pan.pl.
authors.forename: Renata
authors.initials: R
authors.lastname: Plocinska
authors.affiliation: Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland. Electronic address: jdziadek@cbm.pan.pl.
authors.forename: Jarosław
authors.initials: J
authors.lastname: Dziadek
authors.affiliation: Research Institute for Physical Chemical Problems, Belarusian State University, Minsk, Belarus; Faculty of Chemistry, Belarusian State University, Minsk, Belarus. Electronic address: lusova@mail.ru.
authors.forename: Elena
authors.initials: E
authors.lastname: Rudaya
authors.affiliation: Faculty of Chemistry, Belarusian State University, Minsk, Belarus. Electronic address: shadiro@open.by.
authors.forename: Irina
authors.initials: I
authors.lastname: Edimecheva
authors.affiliation: Research Institute for Physical Chemical Problems, Belarusian State University, Minsk, Belarus; Faculty of Chemistry, Belarusian State University, Minsk, Belarus. Electronic address: biopharm@bsu.by.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Shkumatov
mesh_terms: D002784:Cholesterol / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D009159:Mycobacteriaceae / Q000378:metabolism*; D009169:Mycobacterium tuberculosis / Q000378:metabolism; D013256:Steroids / Q000378:metabolism*
chemical_list: D013256:Steroids; D002784:Cholesterol
keywords: 22-NBD-cholesterol (PubChem CID: 127604); 3β-Hydroxysteroid dehydrogenase; Fluorescence microscopy; Fluorescent steroids; Lipid droplets; Mass-spectrometry; Menadione sodium bisulfite (PubChem CID: 23665888); Mycobacteria; Nile Red (PubChem CID: 65182)