Comprehensive proteome analysis of Mycobacterium ulcerans and quantitative comparison of mycolactone biosynthesis.
Mycobacterium ulcerans is the causative agent of Buruli ulcer, a rapidly emerging human disease in which mycolactone, a cytotoxic and immunosuppressive macrocyclic polyketide, is responsible for massive skin destruction. The genome sequencing of M. ulcerans has recently been accomplished (http://genolist.pasteur.fr/BuruList/) enabling the first proteome study of this important human pathogen. Here, we present a comprehensive proteome analysis of different subcellular fractions and culture supernatant of in vitro grown M. ulcerans. By a combination of gel-based and gel-free techniques for protein and peptide separation with subsequent analysis by MS, we identified 1074 different proteins, corresponding to 25% of the protein-coding DNA sequence. Interestingly, new information was obtained about central metabolism and lipid biosynthesis, and as many as 192 conserved hypothetical proteins were found. Comparative analysis of the wild-type strain and an isogenic mycolactone-deficient mutant, by 2-DE and iTRAQ labeling of the cytoplasmic fraction, revealed differences in the expression profiles of proteins involved in lipid metabolism and information pathways, as well as stress responses.
Tafelmeyer
Petra
P
Plate-Forme de Protéomique, Institut Pasteur, Paris, France.
Laurent
Christine
C
Lenormand
Pascal
P
Rousselle
Jean-Claude
JC
Marsollier
Laurent
L
Reysset
Gilles
G
Zhang
Runxuan
R
Sickmann
Albert
A
Stinear
Timothy P
TP
Namane
Abdelkader
A
Cole
Stewart T
ST
D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D001427:Bacterial Toxins / Q000096:biosynthesis*; D015180:Electrophoresis, Gel, Two-Dimensional; D018942:Macrolides; D019911:Mycobacterium ulcerans / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D018942:Macrolides; D020543:Proteome; C117218:mycolactone