QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 6. An APC/C subunit Cut23 is acetylated in vivo. (A) 3HA6His-tagged Cut23 overproduced from pREP1 plasmid in wild type was purified using nickel beads under denaturing conditions and was separated by SDS-PAGE followed by Coomassie Brilliant Blue (CBB) staining. (B) The band of purified Cut23-3HA6His shown in (A) (64 kDa) was cut out and subjected to survey of the acetylation site using LC-MS/MS after trypsin digestion (Ohta et al., 2002). The MS/MS spectra of the tryptic acetylated peptide amino acids 82-95, 123-132, 499-502 of Cut23 obtained by collision-induced dissociation of the [M + 2H]²⁺ or [M + 3H]³⁺ precursor ions, m/z 858.55, m/z 606.87, m/z 496.91, m/z 946.02. Each peptide has one possible acetylated lysine residue, K88 in 82-95, K126 in 123-132, K461 in 453-464 or K502 in 499-514. Peptides containing unmodified lysine residues were also detected for each region and these four acetylated peptides were detected in two independent experiments. (C) Localization of the four putative acetylated sites in Cut23 protein. The acetylation sites (Ac) and TPR domains are shown in the diagram. Six destruction box-like sequences (RxxL) in Cut23 are also shown as black bars. (D) Spot tests for indicated strains on YPD plates in the absence or the presence of 12.5 µg/ml TSA.

Return to article