Publication Information
Xue et al., 2013
No external accession available
Abstract
Mol Cell Proteomics. 2013 Aug;12(8):2354-69. doi: 10.1074/mcp.O113.027284. Epub
2013 May 8.
Quantitative measurement of phosphoproteome response to osmotic stress in
arabidopsis based on Library-Assisted eXtracted Ion Chromatogram (LAXIC).
Xue L(1), Wang P, Wang L, Renzi E, Radivojac P, Tang H, Arnold R, Zhu JK, Tao
WA.
Author information:
(1)Department of Biochemistry, Purdue University, West Lafayette, IN 47907, USA.
Global phosphorylation changes in plants in response to environmental stress
have been relatively poorly characterized to date. Here we introduce a novel
mass spectrometry-based label-free quantitation method that facilitates
systematic profiling plant phosphoproteome changes with high efficiency and
accuracy. This method employs synthetic peptide libraries tailored specifically
as internal standards for complex phosphopeptide samples and accordingly, a
local normalization algorithm, LAXIC, which calculates phosphopeptide abundance
normalized locally with co-eluting library peptides. Normalization was achieved
in a small time frame centered to each phosphopeptide to compensate for the
diverse ion suppression effect across retention time. The label-free LAXIC
method was further treated with a linear regression function to accurately
measure phosphoproteome responses to osmotic stress in Arabidopsis. Among 2027
unique phosphopeptides identified and 1850 quantified phosphopeptides in
Arabidopsis samples, 468 regulated phosphopeptides representing 497 phosphosites
have shown significant changes. Several known and novel components in the
abiotic stress pathway were identified, illustrating the capability of this
method to identify critical signaling events among dynamic and complex
phosphorylation. Further assessment of those regulated proteins may help shed
light on phosphorylation response to osmotic stress in plants.
DOI: 10.1074/mcp.O113.027284
PMCID: PMC3734591
PMID: 23660473 [Indexed for MEDLINE]
