Two-dimensional map of the proteome of Haemophilus influenzae.
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Langen H, Takacs B, Evers S, Berndt P, Lahm HW, Wipf B, Gray C, Fountoulakis M
Two-dimensional map of the proteome of Haemophilus influenzae.
Electrophoresis. 2000 Jan;21(2):411-29.
- PubMed ID
- 10675023 [ View in PubMed]
- 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.