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@INBOOK{Chang:857534,
author = {Bütepage, Mareike and Krieg, Sarah and Eckei, Laura and
Li, Jinyu and Rossetti, Giulia and Verheugd, Patricia and
Lüscher, Bernhard},
editor = {Chang, Paul},
title = {{A}ssessment of {I}ntracellular {A}uto-{M}odification
{L}evels of {ARTD}10 {U}sing
{M}ono-{ADP}-{R}ibose-{S}pecific {M}acrodomains 2 and 3 of
{M}urine {A}rtd8},
volume = {1813},
address = {New York, NY},
publisher = {Springer New York},
reportid = {FZJ-2018-06525},
isbn = {978-1-4939-8587-6 (print)},
series = {Methods in Molecular Biology},
pages = {41 - 63},
year = {2018},
comment = {ADP-ribosylation and NAD+ Utilizing Enzymes / Chang, Paul
(Editor) ; New York, NY : Springer New York, 2018, Chapter 4
; ISSN: 1064-3745=1940-6029 ; ISBN:
978-1-4939-8587-6=978-1-4939-8588-3 ;
doi:10.1007/978-1-4939-8588-3},
booktitle = {ADP-ribosylation and NAD+ Utilizing
Enzymes / Chang, Paul (Editor) ; New
York, NY : Springer New York, 2018,
Chapter 4 ; ISSN: 1064-3745=1940-6029 ;
ISBN:
978-1-4939-8587-6=978-1-4939-8588-3 ;
doi:10.1007/978-1-4939-8588-3},
abstract = {Mono-ADP-ribosylation is a posttranslational modification,
which is catalyzed in cells by certain members of the
ADP-ribosyltransferase diphtheria toxin-like family (ARTD)
of ADP-ribosyltransferases (aka PARP enzymes). It involves
the transfer of a single residue of ADP-ribose (ADPr) from
the cofactor NAD+ onto substrate proteins. Although 12 of
the 17 members of the ARTD family have been defined as
mono-ARTDs in in vitro assays, relatively little is known
about their exact cellular functions. A major challenge is
the detection of mono-ADP-ribosylated (MARylated) proteins
in cells as no antibodies are available that detect
exclusively MARylated proteins. As an alternative to
classical antibodies, the MAR-specific binding domains
macro2 and macro3 of Artd8 can be utilized alone or in
combination, to demonstrate intracellular auto-modification
levels of ARTD10 in cells in both co-immunoprecipitation and
co-localization experiments. Here we demonstrate that
different macrodomain constructs of human ARTD8 and murine
Artd8, alone or in combination, exert differences with
regard to their interaction with ARTD10 in cells. Precisely,
while the macrodomains of murine Artd8 interacted with
ARTD10 in cells in a MARylation-dependent manner, the
macrodomains of human ARTD8 interacted with ARTD10
independent of its catalytic activity. Moreover, we show
that a combination of macro2 and macro3 of murine Artd8 was
recruited more efficiently to ARTD10 during co-localization
experiments compared to the single domains. Therefore,
murine Artd8 macrodomain constructs can serve as a tool to
evaluate intracellular ARTD10 auto-modification levels using
the described methods, while the human ARTD8 macrodomains
are less suited because of ADPr-independent binding to
ARTD10. Protocols for co-immunoprecipitation and
co-localization experiments are described in detail.},
cin = {IAS-5 / JSC},
ddc = {570},
cid = {I:(DE-Juel1)IAS-5-20120330 / I:(DE-Juel1)JSC-20090406},
pnm = {511 - Computational Science and Mathematical Methods
(POF3-511)},
pid = {G:(DE-HGF)POF3-511},
typ = {PUB:(DE-HGF)7},
pubmed = {pmid:30097860},
UT = {WOS:000454740000005},
doi = {10.1007/978-1-4939-8588-3_4},
url = {https://juser.fz-juelich.de/record/857534},
}