An iterative calibration method with prediction of post-translational modifications for the construction of a two-dimensional electrophoresis database of mouse mammary gland proteins

Aksu S., Scheler C., Focks N., Leenders F., Theurıng F., Salnıkow J., ...Daha Fazla

PROTEOMICS, cilt.2, sa.10, ss.1452-1463, 2002 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 2 Sayı: 10
  • Basım Tarihi: 2002
  • Dergi Adı: PROTEOMICS
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.1452-1463
  • Anahtar Kelimeler: breast, database, mammary gland proteins, matrix-assisted laser desorption/ionization, mass spectrometry, two-dimensional gel electrophoresis, calibration, COMPARATIVE PROTEOME ANALYSIS, APOLIPOPROTEIN-A-I, 2-DIMENSIONAL ELECTROPHORESIS, BREAST-CANCER, EXPRESSION, CELLS, IDENTIFICATION, CARCINOMA, CASEIN, TUMOR
  • Akdeniz Üniversitesi Adresli: Evet

Özet

Protein databases serve as general reference recources 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 (p 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 Mr and p/ 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.

Protein databases serve as general reference recources 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 (p 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 Mr and p/ 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.