Two-dimensional map of the proteome of Haemophilus influenzae.
We have constructed a two-dimensional database of the proteome of Haemophilus influenzae, a bacterium of medical interest of which the complete genome, comprising about 1742 open reading frames, has been sequenced. The soluble protein fraction of the microorganism was analyzed by two-dimensional electrophoresis, using immobilized pH gradient strips of various pH regions, gels with different acrylamide concentrations and buffers with different trailing ions. In order to visualize low-copy-number gene products, we employed a series of protein extraction and sample application approaches and several chromatographic steps, including heparin chromatography, chromatofocusing and hydrophobic interaction chromatography. We have also analyzed the cell envelope-bound protein fraction using either immobilized pH gradient strips or a two-detergent system with a cationic detergent in the first and an anionic detergent in the second-dimensional separation. Different proteins (502) were identified by matrix-assisted laser desorption/ionization mass spectrometry and amino acid composition analysis. This is at present one of the largest two-dimensional proteome databases.
Langen
H
H
Genomics Technologies, F. Hoffmann-La Roche Ltd., Pharmaceutical Research, Basel, Switzerland.
Takács
B
B
Evers
S
S
Berndt
P
P
Lahm
H W
HW
Wipf
B
B
Gray
C
C
Fountoulakis
M
M
D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006193:Haemophilus influenzae; D020543:Proteome / Q000032:analysis*
D020543:Proteome