原创力文档artd10 substrate identification on protein microarrays regulation of gsk3beta by mono-adp-ribosylationartd10底物识别蛋白质微阵列的监管gsk3beta mono-adp-ribosylation.pdfVIP
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artd10 substrate identification on protein microarrays regulation of gsk3beta by mono-adp-ribosylationartd10底物识别蛋白质微阵列的监管gsk3beta mono-adp-ribosylation
Feijs et al. Cell Communication and Signaling 2013, 11:5
/content/11/1/5
RESEARCH Open Access
ARTD10 substrate identification on protein
microarrays: regulation of GSK3β by
mono-ADP-ribosylation
1 1,3 1,4 1 1 1
Karla LH Feijs , Henning Kleine , Anne Braczynski , Alexandra H Forst , Nicolas Herzog , Patricia Verheugd ,
Ulrike Linzen1,5, Elisabeth Kremmer2 and Bernhard Lüscher1*
Abstract
Background: Although ADP-ribosylation has been described five decades ago, only recently a distinction has been
made between eukaryotic intracellular poly- and mono-ADP-ribosylating enzymes. Poly-ADP-ribosylation by ARTD1
(formerly PARP1) is best known for its role in DNA damage repair. Other polymer forming enzymes are ARTD2
(formerly PARP2), ARTD3 (formerly PARP3) and ARTD5/6 (formerly Tankyrase 1/2), the latter being involved in Wnt
signaling and regulation of 3BP2. Thus several different functions of poly-ADP-ribosylation have been well described
whereas intracellular mono-ADP-ribosylation is currently largely undefined. It is for example not known which
proteins function as substrate for the different mono-ARTDs. This is partially due to lack of suitable reagents to
study mono-ADP-ribosylation, which limits the current understanding of this post-translational modification.
Results: We have optimized a novel screening method employing protein microarrays, ProtoArrays®, applied here
for the identification of substrates of ARTD10 (formerly PARP10) and ARTD8 (formerly PARP14). The results of this
substrate screen were validated using in vitro ADP-ribosylation assays with recombinant proteins. Further analysis of
the novel ARTD10 substrate GSK3β revealed mono-ADP-ribosylation as a regulatory mechan
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