Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2009 Sep 22;37(21):7072–7084. doi: 10.1093/nar/gkp777

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2009. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 6. — Cdk2 is a substrate acetylated by PCAF in vitro. (A) Purified GST-cdk2 was subjected to in vitro acetylation assays using the catalytic domain of PCAF [GST-HAT(PCAF)], GST-CBP or GST-Tip60 in the presence of [¹⁴C]acetylCoA. Purified GST was used as a negative control substrate. In the assays with PCAF or Tip60 their autoacetylation was used as a positive control, whereas in the case of CBP, histones were used as a positive control substrate (see Supplementary Data Figure S4A). Acetylated proteins were visualized by autoradiography (top panel). A loading control gel was stained with coomassie blue (bottom panel). (B) Purified recombinant cdk2 fragments were subjected to in vitro acetylation assays as in (A) using GST-HAT (PCAF) as acetylase. Acetylated proteins were visualized by autoradiography (top panel). A loading control gel was stained with coomassie blue (bottom panel). (C) Fourteen peptides including one or two consecutive lysines from the cdk2 fragment including aa 1–106, were spotted on a membrane. As a positive control a peptide from histone H3 was added. The membrane was subjected to in vitro acetylation assays with GST-HAT(PCAF) and [¹⁴C]acetylCoA. Acetylation was visualized by autoradiography. (D) Purified GST-cdk2 WT and GST-cdk2 K33R were subjected to in vitro acetylation assays with GST-HAT(PCAF). Acetylation was visualized by autoradiography (top panel). A loading control gel was stained with red ponceau (bottom panel).