A purification strategy for analysis of the DNA/RNA-associated sub-proteome from chloroplasts of mustard cotyledons

摘要: Plant cotyledons are a tissue that is particularly active in plastid gene expression order to develop functional chloroplasts from pro-plastids, the precursor stage plant embryos. Cotyledons, therefore, represent material being ideal for study of composition, function and regulation protein complexes involved expression. Here, we present pilot uses heparin-Sepharose phospho-cellulose chromatography combination with isoelectric focussing denaturing SDS gel electrophoresis (two-dimensional electrophoresis) investigating nucleic acids binding sub-proteome mustard purified cotyledons. We describe technical requirements highly resolved biochemical purification several hundreds spots obtained such samples. Subsequent mass spectrometry peptides isolated out cut had been treated trypsin identified 58 different proteins within 180 distinct spots. Our analyses indicate high enrichment transcription translation and, addition, presence massive post-translational modification this sub-fraction. The provides an extended catalog its describes suitable strategy further analysis low abundant related proteins.