title: An iterative calibration method with prediction of post-translational modifications for the construction of a two-dimensional electrophoresis database of mouse mammary gland proteins.
abstract: Protein databases serve as general reference resources providing an orientation on two-dimensional electrophoresis (2-DE) patterns of interest. The intention behind constructing a 2-DE database of the water soluble proteins from wild-type mouse mammary gland tissue was to create a reference before going on to investigate cancer-associated protein variations. This database shall be deemed to be a model system for mouse tissue, which is open for transgenic or knockout experiments. Proteins were separated and characterized in terms of their molecular weight (M(r)) and isoelectric point (pI) by high resolution 2-DE. The proteins were identified using prevalent proteomics methods. One method was peptide mass fingerprinting by matrix-assisted laser desorption/ionization-mass spectrometry. Another method was N-terminal sequencing by Edman degradation. By N-terminal sequencing M(r) and pI values were specified more accurately and so the calibration of the master gel was obtained more systematically and exactly. This permits the prediction of possible post-translational modifications of some proteins. The mouse mammary gland 2-DE protein database created presently contains 66 identified protein spots, which are clickable on the gel pattern. This relational database is accessible on the WWW under the URL: http://www.mpiib-berlin.mpg.de/2D-PAGE.
authors.affiliation: Technical University, Max-Volmer Institute for Biophysical Chemistry and Biochemistry, Berlin, Germany.
authors.forename: Sevil
authors.initials: S
authors.lastname: Aksu
authors.forename: Christian
authors.initials: C
authors.lastname: Scheler
authors.forename: Nicole
authors.initials: N
authors.lastname: Focks
authors.forename: Frauke
authors.initials: F
authors.lastname: Leenders
authors.forename: Franz
authors.initials: F
authors.lastname: Theuring
authors.forename: Johann
authors.initials: J
authors.lastname: Salnikow
authors.forename: Peter R
authors.initials: PR
authors.lastname: Jungblut
mesh_terms: D000818:Animals; D002138:Calibration; D019992:Databases as Topic; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005260:Female; D006863:Hydrogen-Ion Concentration; D008321:Mammary Glands, Animal / Q000378:metabolism*; D051379:Mice; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D020543:Proteome