title: '98 Escherichia coli SWISS-2DPAGE database update.
abstract: The combination of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), computer image analysis and several protein identification techniques allowed the Escherichia coli SWISS-2DPAGE database to be established. This is part of the ExPASy molecular biology server accessible through the WWW at the URL address http://www.expasy.ch/ch2d/ch2d-top.html . Here we report recent progress in the development of the E. coli SWISS-2DPAGE database. Proteins were separated with immobilized pH gradients in the first dimension and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. To increase the resolution of the separation and thus the number of identified proteins, a variety of wide and narrow range immobilized pH gradients were used in the first dimension. Micropreparative gels were electroblotted onto polyvinylidene difluoride membranes and spots were visualized by amido black staining. Protein identification techniques such as amino acid composition analysis, gel comparison and microsequencing were used, as well as a recently described Edman "sequence tag" approach. Some of the above identification techniques were coupled with database searching tools. Currently 231 polypeptides are identified on the E. coli SWISS-2DPAGE map: 64 have been identified by N-terminal microsequencing, 39 by amino acid composition, and 82 by sequence tag. Of 153 proteins putatively identified by gel comparison, 65 have been confirmed. Many proteins have been identified using more than one technique. Faster progress in the E. coli proteome project will now be possible with advances in biochemical methodology and with the completion of the entire E. coli genome.
authors.affiliation: Clinical Chemistry Laboratory, Geneva University Hospital, Switzerland. luisa.tonella@dim.hcuge.ch
authors.forename: L
authors.initials: L
authors.lastname: Tonella
authors.forename: B J
authors.initials: BJ
authors.lastname: Walsh
authors.forename: J C
authors.initials: JC
authors.lastname: Sanchez
authors.forename: K
authors.initials: K
authors.lastname: Ou
authors.forename: M R
authors.initials: MR
authors.lastname: Wilkins
authors.forename: M
authors.initials: M
authors.lastname: Tyler
authors.forename: S
authors.initials: S
authors.lastname: Frutiger
authors.forename: A A
authors.initials: AA
authors.lastname: Gooley
authors.forename: I
authors.initials: I
authors.lastname: Pescaru
authors.forename: R D
authors.initials: RD
authors.lastname: Appel
authors.forename: J X
authors.initials: JX
authors.lastname: Yan
authors.forename: A
authors.initials: A
authors.lastname: Bairoch
authors.forename: C
authors.initials: C
authors.lastname: Hoogland
authors.forename: F S
authors.initials: FS
authors.lastname: Morch
authors.forename: G J
authors.initials: GJ
authors.lastname: Hughes
authors.forename: K L
authors.initials: KL
authors.lastname: Williams
authors.forename: D F
authors.initials: DF
authors.lastname: Hochstrasser
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis*; D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000737:chemistry*; D007526:Isoelectric Point; D008567:Membranes, Artificial; D011145:Polyvinyls; D013194:Staining and Labeling / Q000379:methods; D005465:Fluorocarbon Polymers
chemical_list: D001426:Bacterial Proteins; D008567:Membranes, Artificial; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers