Continued proteomic analysis of Mycobacterium leprae subcellular fractions.
Recently the sequence of the Mycobacterium leprae chromosome, the only known obligate intracellular mycobacterium, was completed. It has a dramatic reduction in functional genes, with a coding capacity of only 49.5%, the lowest one so far observed among bacterial genomes. The leprosy bacillus seems to preserve a minimal set of genes that allows its survival in the host. The identification of genes that are actually expressed by the bacterium is of high significance in the context of mycobacterial pathogenesis. In this current study, a proteomic approach was undertaken to identify the proteins present in the soluble/cytosol and membrane subcellular fractions obtained from armadillo derived M. leprae. Proteins from each fraction were separated by two-dimensional gel electrophoresis (2-DE) and identified by mass spectrometry. A total of 147 protein spots were identified from 2-DE patterns and shown to comprise products of 44 different genes, twenty eight of them corresponding to new proteins. Additionally, two highly basic proteins (with pI >10.0) were isolated by heparin affinity chromatography and identified by N-terminal sequencing. This study constitutes the first application of proteomics to a host-derived Mycobacterium.
Marques
Maria Angela M
MA
Department of Microbiology, Colorado State University, Fort Collins, CO 80523-1682, USA.
Espinosa
Benjamin J
BJ
Xavier da Silveira
Erika K
EK
Pessolani
Maria Cristina V
MC
Chapeaurouge
Alex
A
Perales
Jonas
J
Dobos
Karen M
KM
Belisle
John T
JT
Spencer
John S
JS
Brennan
Patrick J
PJ
D001426:Bacterial Proteins / Q000737:chemistry*; D002846:Chromatography, Affinity; D003600:Cytosol / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D016680:Genome, Bacterial; D013058:Mass Spectrometry; D009166:Mycobacterium leprae / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization; D013347:Subcellular Fractions / Q000378:metabolism
D001426:Bacterial Proteins; D020543:Proteome