Peptide mass fingerprinting of chaperonin‐containing TCP‐1 (CCT) and copurifying proteins

The chaperonin‐containing TCP‐1 (CCT), found in the eukaryotic cytosol, is currently the focus of extensive research. CCT isolated from mouse testis lysate sediments at 20S in a sucrose gradient and accounts for about 70% of the total protein in this fraction. We intend to identify all the other proteins that copurify with CCT and to compile a reference profile for future studies. Their identification can be accelerated by a combination of protease digestion, matrix‐assisted laser desorption‐mass spectrometry, and database matching known as peptide mass fingerprinting. We applied this strategy to 32 polypeptides resolved by 2‐dimensional gel electrophoresis, and 23 known proteins and 6 novel proteins were identified. We analyzed isoelectric variants of the CCT subunits and differences in the peptide mass spectra of two CCTθ isoforms indicated a novel posttranslational modification of this subunit.—Hynes, G., Sutton, C. W., U, S., Willison, K. R. Peptide mass fingerprinting of chaperonin‐ containing TCP‐l (CCT) and copurifying proteins. FASEB J. 10, 137‐147 (1996)

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