Photoaffinity labelling with the cytokinin agonist azido-CPPU of a 34 kDa peptide of the intracellular pathogenesis-related protein family in the moss Physcomitrella patens.

摘要: As in higher plants, the development of moss Physcomitrella patens is regulated by environmental signals and phytohormones. At protonema level transition from chloronema to caulonema cells under auxin control. The formation on second sub-apical buds that will give rise leafy gametophore requires cytokinins. Using [3H]azidoCPPU (1-(2-azido-6-chloropyrid-4-yl)-3-(4-[3H])phenylurea), a photoactivatable cytokinin agonist, we have specifically photolabelled soluble 34 kDa protein P. patens. Urea derivatives were very efficient competitors photolabelling while purine-type cytokinins poor competitors. UBP34 was purified affinity chromatography sequences six internal peptides obtained. A cDNA encoding cloned screening library with probe amplified PCR using degenerate primers designed peptide sequences. amino acid sequence shows an average identity 42% both intracellular PR proteins BetV1-related family plant allergens. Recombinant expressed Escherichia coli confirmed bind azidoCPPU.