Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection.
Capillary electrophoresis (CE) coupled with laser-induced fluorescence detection (LIF) has allowed to obtain protein fingerprints, which have demonstrated to be useful in microorganisms characterization. In this work, protein fingerprints of two species of Staphylococcus grown in different culture media and submitted to temperature and nitrosative stress were studied by CE-LIF. After the growth of the bacteria, protein extracts were obtained by cell lysis using sonication. The water-soluble fraction of these lysates was derivatized on-capillary with a fluorogenic dye, 3-(2-furoyl)quinoline-2-carboxaldehyde. The fluorescent products were analyzed by CE using phosphate buffer containing submicellar concentrations of sodium pentanesulfate and detected by LIF. Different protein fingerprints were obtained depending on the bacterial species studied, indicating the usefulness of this method for the identification of different species of the same bacterial genus. It was also demonstrated that the CE protein fingerprints were dependent on the culture conditions, such as growth medium, or on stressing conditions, such as heat shock or nitrosative stress.
Veledo
Maria Teresa
MT
Instituto de Química Orgánica General (CSIC), Juan de la Cierva 3, 28006 Madrid, Spain.
Pelaez-Lorenzo
Cristina
C
Gonzalez
Ramon
R
de Frutos
Mercedes
M
Diez-Masa
Jose Carlos
JC
D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D005663:Furans / Q000737:chemistry*; D007834:Lasers; D018384:Oxidative Stress; D010449:Peptide Mapping / Q000379:methods*; D011804:Quinolines / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013210:Staphylococcus / Q000378:metabolism*; D013696:Temperature
D005456:Fluorescent Dyes; D005663:Furans; D011804:Quinolines; C063756:3-(2-furoyl)quinoline-2-carbaldehyde