Titanium Oxide-Based Phosphopeptide Enrichment from Arabidopsis Seedlings.

Protein phosphorylation is one of the most widely studied posttranslational modifications, and its role in signal transduction has gained particular attention. The relatively low abundance of the phosphorylated form of proteins makes identification by mass spectrometry challenging in the absence of selective enrichment. Titanium oxide-based enrichment of phosphopeptides in the presence of acidic modifiers is highly selective and makes it technically possible to detect thousands of phosphopeptides in a single sample by liquid chromatography-mass spectrometry.

Regulation of pattern recognition receptor signalling by phosphorylation and ubiquitination.

Our understanding of how plant innate immunity is triggered and regulated has seen tremendous progress over the last decade, with many important players identified in the model systems Arabidopsis thaliana and Oryza sativa (rice). Identification of these components has come from both genetic screens as well as from proteomics approaches. While genetic approaches are powerful tools of discovery to identify key components in a signalling pathway, the application of genetics is limited when dealing with redundancy or when mutations cause lethal phenotypes.

Attenuation of pattern recognition receptor signaling is mediated by a MAP kinase kinase kinase.

Pattern recognition receptors (PRRs) play a key role in plant and animal innate immunity. PRR binding of their cognate ligand triggers a signaling network and activates an immune response. Activation of PRR signaling must be controlled prior to ligand binding to prevent spurious signaling and immune activation.

Phosphopeptide immuno-affinity enrichment to enhance detection of tyrosine phosphorylation in plants.

Tyrosine (Tyr) phosphorylation plays an essential role in signaling in animal systems, but the relative contribution of Tyr phosphorylation to plant signal transduction has, until recently, remained an open question. One of the major issues hampering the analysis is the low abundance of Tyr phosphorylation and therefore underrepresentation in most mass spec-based proteomic studies. Here, we describe a working approach to selectively enrich Tyr-phosphorylated peptides from complex plant tissue samples.

Targeted quantitative phosphoproteomics approach for the detection of phospho-tyrosine signaling in plants.

Tyrosine (Tyr) phosphorylation plays an essential role in signaling in animal systems. However, a few studies have also reported Tyr phosphorylation in plants, but the relative contribution of tyrosine phosphorylation to plant signal transduction has remained an open question. We present an approach to selectively measure and quantify Tyr phosphorylation in plant cells, which can also be applied to whole plants.

Phosphoproteomics perspective on plant signal transduction and tyrosine phosphorylation.

Plants and animal cells use intricate signaling pathways to respond to a diverse array of stimuli. These stimuli include signals from environment, such as biotic and abiotic stress signals, as well as cell-to-cell signaling required for pattern formation during development. The transduction of the signal often relies on the post-translational modification (PTM) of proteins.

Listeria monocytogenes sigma B regulates stress response and virulence functions.

While the stress-responsive alternative sigma factor sigma(B) has been identified in different species of Bacillus, Listeria, and Staphylococcus, the sigma(B) regulon has been extensively characterized only in B. subtilis. We combined biocomputing and microarray-based strategies to identify sigma(B)-dependent genes in the facultative intracellular pathogen Listeria monocytogenes.