Similarly, sumoylation of PML directs p53 to PML NBs and could then trigger some modification, such as acetylation and sumoylation, which stimulates the transcriptional activity of p Lysate was reduced by 4. Previous studies indicated that SUMO modification could be stimulated by cell stresses such as heat shock and proteasome inhibition 30 The same population of cells or tissue can be subjected to analysis of both Ub attachment and SUMO attachment simply by splitting the sample in two and digesting half with trypsin and the other half with WaLP. Meyer JG, et al. Hang, J. Ahmad1 Karl R.
For example, Hendriks et al. introduced a lysine-deficient SUMO-3 with . then Hela cells were washed with cold PBS, and harvested with 8 M. SUMO proteins are covalently attached to lysine residues of proteins, which are.
The BTBContaining Protein Kctd15 Is SUMOylated In Vivo
on ice, remove media by aspiration and add 1ml of ice-cold PBS to the plate. We found % of all SUMOylated lysines to reside in the KxE-type Lyophilized peptides were dissolved in ice-cold SUMO-IP Buffer (
The lysate can be simply split in half, with one half digested with WaLP and the other half digested with trypsin Fig. Rangasamy, D. Using the ubiquitin-modified proteome to monitor protein homeostasis function.
The peptides were eluted in two steps. Meyer1 Eric J.
The ubiquitin-like protein SUMO is a regulator involved in most cellular mechanisms. as cold-resistant MT bundles called kinetochore-fibers (k-fibers) .
Conjugation of SUMO to a protein occupies a lysine residue and. SUMO is attached to a Lysine contained in a tetrapeptide motif with the . Cells were lysed in cold lysis buffer (% Triton X, 50 mM.
Eifler K, Vertegaal AC. The mechanism involved in maturation and transfer of SUMO to target substrates is very similar to that seen with ubiquitination and other ubiquitin-like proteins 34.
Stade, K. Lysines 11 and 48 on Nedd8 were observed to be modified by both Ub and SUMO and the extent of modification was largely unperturbed by proteasome inhibition Fig. An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database.