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@ARTICLE{Arsova:851057,
author = {Arsova, Borjana and Watt, Michelle and Usadel, Björn},
title = {{M}onitoring of {P}lant {P}rotein {P}ost-translational
{M}odifications {U}sing {T}argeted {P}roteomics},
journal = {Frontiers in plant science},
volume = {Vol. 9},
issn = {1664-462X},
address = {Lausanne},
publisher = {Frontiers Media},
reportid = {FZJ-2018-04770},
pages = {1168},
year = {2018},
abstract = {Protein posttranslational modifications (PTMs) are among
the fastest and earliest of plant responses to changes in
the environment, making the mechanisms and dynamics of PTMs
an important area of plant science. One of the most studied
PTMs is protein phosphorylation. This review summarizes the
use of targeted proteomics for the elucidation of the
biological functioning of plant PTMs, and focuses primarily
on phosphorylation. Since phosphorylated peptides have a low
abundance, usually complex enrichment protocols are required
for their research. Initial identification is usually
performed with discovery phosphoproteomics, using high
sensitivity mass spectrometers, where as many
phosphopeptides are measured as possible. Once a PTM site is
identified, biological characterization can be addressed
with targeted proteomics. In targeted proteomics,
Selected/Multiple Reaction Monitoring (S/MRM) is
traditionally coupled to simple, standard protein digestion
protocols, often omitting the enrichment step, and relying
on triple-quadruple mass spectrometer. The use of synthetic
peptides as internal standards allows accurate
identification, avoiding cross-reactivity typical for some
antibody based approaches. Importantly, internal standards
allow absolute peptide quantitation, reported down to 0.1
femtomoles, also useful for determination of phospho-site
occupancy. S/MRM is advantageous in situations where
monitoring and diagnostics of peptide PTM status is needed
for many samples, as it has faster sample processing times,
higher throughput than other approaches, and excellent
quantitation and reproducibility. Furthermore, the number of
publicly available data-bases with plant PTM discovery data
is growing, facilitating selection of modified peptides and
design of targeted proteomics workflows. Recent instrument
developments result in faster scanning times, inclusion of
ion-trap instruments leading to parallel reaction
monitoring- which further facilitates S/MRM experimental
design. Finally, recent combination of data independent and
data dependent spectra acquisition means that in addition to
anticipated targeted data, spectra can now be queried for
unanticipated information. The potential for future
applications in plant biology is outlined.},
cin = {IBG-2},
ddc = {570},
cid = {I:(DE-Juel1)IBG-2-20101118},
pnm = {582 - Plant Science (POF3-582)},
pid = {G:(DE-HGF)POF3-582},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30174677},
UT = {WOS:000441883100001},
doi = {10.3389/fpls.2018.01168},
url = {https://juser.fz-juelich.de/record/851057},
}