Mass spectrometric characterization of mitochondrial complex I NDUFA10 variants.

In the present study, we have used a combination of 2-DE and MS to isolate and characterize two variants of the mitochondrial complex I subunit NDUFA10 from Wistar rat brain. Extensive MS/MS analysis revealed that a D/N substitution at position 120 resulting from a 353A/G transition in the coding gene is the biochemical difference between the two most abundant NDUFA10 isoforms. Moreover, 33 modifications of distinct chemical nature targeting 59 specific residues were found to be common to the acidic and basic forms.

Detection of artifacts and peptide modifications in liquid chromatography/mass spectrometry data using two-dimensional signal intensity map data visualization.

We demonstrate how visualization of liquid chromatography/mass spectrometry data as a two-dimensional signal intensity map can be used to assess the overall quality of the data, for the identification of polymer contaminants and artifacts, as well as for the confirmation of post-translational modifications.

Reproducibility of LC-MS-based protein identification.

Traditional analysis of liquid chromatography-mass spectrometry (LC-MS) data, typically performed by reviewing chromatograms and the corresponding mass spectra, is both time-consuming and difficult. Detailed data analysis is therefore often omitted in proteomics applications. When analysing multiple proteomics samples, it is usually only the final list of identified proteins that is reviewed.

Photochromic chromopeptides derived from phycoerythrocyanin: biophysical and biochemical characterization.

Truncated chromopeptides have been prepared from the small photo- and redox-switchable biliprotein alpha-phycoerythrocyanin (alpha-PEC). The native chromoprotein consists of a C-terminal globin domain containing the chromophore and the regulatory cysteins 98 and 99, and a two-helix (X,Y) N-terminal domain responsible for aggregation. Digestion with chymotrypsin-free trypsin leads to three chromopeptides, (N-30, N-33 and N-35), basically lacking the two N-terminal helices X and Y.

Purification, crystallization, NMR spectroscopy and biochemical analyses of alpha-phycoerythrocyanin peptides.

The alpha-phycoerythrocyanin subunits of the different phycoerythrocyanin complexes of the phycobilisomes from the cyanobacterium Mastigocladus laminosus perform a remarkable photochemistry. Similar to phytochromes - the photoreceptors of higher plants - the spectral properties of the molecule reversibly change according to the irradiation wavelength. To enable extensive analyses, the protein has been produced at high yield by improving purification protocols.