Proteome cataloging and relative quantification of Francisella tularensis tularensis strain Schu4 in 2D PAGE using preparative isoelectric focusing.
The protein complement of whole cell extract of the bacterium Francisella tularensis tularensis was analyzed using two-dimensional electrophoresis with preparative isoelectric focusing in the first dimension. The format allows the quantification of relative protein abundance by linear densitometry and extends the potential dynamic range of protein detection by as much as an order of magnitude. The relative abundance and rank order of 136 unique proteins identified in F. tularensis tularensis were established. It is estimated that 16% of the moderately to highly expressed proteins and 8% of all predicted non-pseudogenes were identified by comparing this proteome information with the relative abundance of mRNA as measured by microarray. This rank-ordered proteome list provides an important resource for understanding the pathogenesis of F. tularensis and is a tool for the selection and design of synthetic vaccines. This method represents a useful additional technique to improve whole proteome analyses of simple organisms.
University of Tennessee Health Science Center, Center of Excellence in Genomics and Bioinformatics, Department of Molecular Sciences, Memphis, Tennessee 38163, USA.
Robert H
RH
Waldo
Edwin D
ED
Cummings
Siva T
ST
Sarva
Janet M
JM
Brown
Crystal M
CM
Lauriano
Lorne A
LA
Rose
Robert J
RJ
Belland
Karl E
KE
Klose
George M
GM
Hilliard
D001426:Bacterial Proteins / Q000737:chemistry; D019295:Computational Biology / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005604:Francisella tularensis / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods*; D040081:Protein Array Analysis / Q000379:methods*; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism; D016622:Silver Staining
D001426:Bacterial Proteins; D020543:Proteome; D012333:RNA, Messenger