Proteomics

Dataset Information

0

A novel tool, Multiple Marker Abundance Profiling, uses Selected Reaction Monitoring and data-dependent acquisition for rapid estimation of organelle abundance in subcellular samples.


ABSTRACT: Methods were investigated that would deliver rapid and semi-quantitative analysis of the subcellular composition of Arabidopsis samples, as the standard technique for this i.e. immunoblotting is difficult to quantify and relies on a very few protein targets, whose responses to developmental or environmental cues are often not known. Spectral counts from shotgun analysis of different Arabidopsis tissues, cells or growth stages were compared to selected reaction monitoring (SRM) analyses of the same samples. Results were further compared to a novel protein abundance scoring method, MASCP Gator scoring, described in this study for the first time. The latter is reliant on previously collated public proteomics data and so provides an estimate of protein abundance even in the absence of de-novo experimentation. This scoring method is presented as an online tool, Multiple Marker Abundance Profiling (MMAP), available at http:SUBA.live. This submission comprises the shotgun data from this study. SRM results are available in a separate submission: http://www.peptideatlas.org/PASS/PASS00906

INSTRUMENT(S): TripleTOF 5600, Q Exactive

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Plant Cell, Cell Suspension Culture, Root, Photosynthetic Cell, Leaf, Rosette, Shoot

SUBMITTER: Harriet Parsons  

LAB HEAD: Harriet Parsons

PROVIDER: PXD005408 | Pride | 2018-10-26

REPOSITORIES: Pride

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Publications

Multiple marker abundance profiling: combining selected reaction monitoring and data-dependent acquisition for rapid estimation of organelle abundance in subcellular samples.

Hooper Cornelia M CM   Stevens Tim J TJ   Saukkonen Anna A   Castleden Ian R IR   Singh Pragya P   Mann Gregory W GW   Fabre Bertrand B   Ito Jun J   Deery Michael J MJ   Lilley Kathryn S KS   Petzold Christopher J CJ   Millar A Harvey AH   Heazlewood Joshua L JL   Parsons Harriet T HT  

The Plant journal : for cell and molecular biology 20171120 6


Measuring changes in protein or organelle abundance in the cell is an essential, but challenging aspect of cell biology. Frequently-used methods for determining organelle abundance typically rely on detection of a very few marker proteins, so are unsatisfactory. In silico estimates of protein abundances from publicly available protein spectra can provide useful standard abundance values but contain only data from tissue proteomes, and are not coupled to organelle localization data. A new protein  ...[more]

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