title: Two-dimensional map of the proteome of Haemophilus influenzae.
abstract: 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.
authors.affiliation: Genomics Technologies, F. Hoffmann-La Roche Ltd., Pharmaceutical Research, Basel, Switzerland.
authors.forename: H
authors.initials: H
authors.lastname: Langen
authors.forename: B
authors.initials: B
authors.lastname: Takács
authors.forename: S
authors.initials: S
authors.lastname: Evers
authors.forename: P
authors.initials: P
authors.lastname: Berndt
authors.forename: H W
authors.initials: HW
authors.lastname: Lahm
authors.forename: B
authors.initials: B
authors.lastname: Wipf
authors.forename: C
authors.initials: C
authors.lastname: Gray
authors.forename: M
authors.initials: M
authors.lastname: Fountoulakis
mesh_terms: D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006193:Haemophilus influenzae; D020543:Proteome / Q000032:analysis*
chemical_list: D020543:Proteome