Two-dimensional gel database of human breast carcinoma cell expressed proteins: An update
- Author(s)
- Rasmussen, RK; Ji, H; Eddes, JS; Zugaro, LM; Reid, GE; Simpson, RJ; Dorow, DS;
- Details
- Publication Year 1998-05,Volume 19,Issue #5,Page 818-825
- Journal Title
- ELECTROPHORESIS
- Publication Type
- Journal Article
- Abstract
- Previously, we reported a two-dimensional gel map and database with molecular weight/isoelectric point (M-r/pI) loci for 22 proteins expressed in the breast carcinoma cell line, MDA-MB231 (Rasmussen et al., Electrophoresis 1997, 18, 588-598). Here we update this database with M-r/pI loci for a further nine cytoplasmic proteins and three Triton X-114 solubilised membrane proteins from MDA-MB231 cells. In addition, a novel protein, previously represented only in expressed sequence tag (EST) databases, has been identified as a Triton X-114 soluble protein and assigned an M-r/pI locus. During the course of isolating proteins from the Triton X-114 fraction, we compared recoveries of proteins in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gels after isoelectric focusing (IEF) using either immobilised pH gradients or carrier ampholytes. In these experiments, a significantly higher proportion of membrane proteins were visible in SDS-polyacrylamide gels after the use of carrier ampholytes for the first dimension. We also report our mass spectrometric-based procedure for identifying two-dimensional electrophoresis (2-DE) gel-resolved proteins, combining in-gel enzymatic digestion, 0.2 mm internal diameter (ID) capillary column reversed-phase high-performance liquid chromatography (RP-HPLC) peptide mapping and electrospray ionisation - ion trap - mass spectrometry.
- Publisher
- WILEY-V C H VERLAG GMBH
- Keywords
- GROWTH-FACTOR RECEPTOR; AMINO-ACID-SEQUENCES; GTP-BINDING PROTEINS; MOLECULAR-CLONING; ELECTROPHORETIC ANALYSIS; MASS-SPECTROMETRY; CANCER; PEPTIDES; IDENTIFICATION; LYMPHOCYTES
- Publisher's Version
- https://doi.org/10.1002/elps.1150190536
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 1998-05-01 12:00:00