title: Identification of proteins induced by polycyclic aromatic hydrocarbon in Mycobacterium vanbaalenii PYR-1 using two-dimensional polyacrylamide gel electrophoresis and de novo sequencing methods.
abstract: Protein profiles of Mycobacterium vanbaalenii PYR-1 grown in the presence of high-molecular-weight polycyclic aromatic hydrocarbons (HMW PAHs) were examined by two-dimensional gel electrophoresis (2-DE). Cultures of M. vanbaalenii PYR-1 were incubated with pyrene, pyrene-4,5-quinone (PQ), phenanthrene, anthracene, and fluoranthene. Soluble cellular protein fractions were analyzed and compared, using immobilized pH gradient (IPG) strips. More than 1000 gel-separated proteins were detected using a 2-DE analysis program within the window of isoelectric point (pI) 4-7 and a molecular mass range of 10-100 kDa. We observed variations in the protein composition showing the upregulation of multiple proteins for the five PAH treatments compared with the uninduced control sample. By N-terminal sequencing or mass spectrometry, we further analyzed the proteins separated by 2-DE. Due to the lack of genome sequence information for this species, protein identification provided an analytical challenge. Several PAH-induced proteins were identified including a catalase-peroxidase, a putative monooxygenase, a dioxygenase small subunit, a small subunit of naphthalene-inducible dioxygenase, and aldehyde dehydrogenase. We also identified proteins related to carbohydrate metabolism (enolase, 6-phosphogluconate dehydrogenase, indole-3-glycerol phosphate synthase, and fumarase), DNA translation (probable elongation factor Tsf), heat shock proteins, and energy production (ATP synthase). Many proteins from M. vanbaalenii PYR-1 showed similarity with protein sequences from M. tuberculosis and M. leprae. Some proteins were detected uniquely upon exposure to a specific PAH whereas others were common to more than one PAH, which indicates that induction triggers not only specific responses but a common response in this strain.
authors.affiliation: Division of Microbiology, National Center for Toxicological Research/U.S. FDA, Jefferson, AR 72079, USA.
authors.forename: Seong-Jae
authors.initials: SJ
authors.lastname: Kim
authors.forename: Richard C
authors.initials: RC
authors.lastname: Jones
authors.forename: Chang-Jun
authors.initials: CJ
authors.lastname: Cha
authors.forename: Ohgew
authors.initials: O
authors.lastname: Kweon
authors.forename: Ricky D
authors.initials: RD
authors.lastname: Edmondson
authors.forename: Carl E
authors.initials: CE
authors.lastname: Cerniglia
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D016678:Genome; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007525:Isoelectric Focusing; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000378:metabolism*; D011084:Polycyclic Aromatic Hydrocarbons / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000032:analysis / Q000737:chemistry; D006180:Proton-Translocating ATPases / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D007477:Ions; D011084:Polycyclic Aromatic Hydrocarbons; D011506:Proteins; D006180:Proton-Translocating ATPases