title: Biochemical and structural properties of cyanases from Arabidopsis thaliana and Oryza sativa.
abstract: Cyanate is toxic to all organisms. Cyanase converts cyanate to CO₂ and NH₃ in a bicarbonate-dependent reaction. The biophysical functions and biochemical characteristics of plant cyanases are poorly studied, although it has been investigated in a variety of proteobacteria, cyanobacteria and fungi. In this study, we characterised plant cyanases from Arabidopsis thaliana and Oryza sativa (AtCYN and OsCYN). Prokaryotic-expressed AtCYN and OsCYN both showed cyanase activity in vitro. Temperature had a similar influence on the activity of both cyanases, but pH had a differential impact on AtCYN and OsCYN activity. Homology modelling provided models of monomers of AtCYN and OsCYN, and a coimmunoprecipitation assay and gel filtration indicated that AtCYN and OsCYN formed homodecamers. The analysis of single-residue mutants of AtCYN indicated that the conserved catalytic residues also contributed to the stability of the homodecamer. KCNO treatment inhibited Arabidopsis germination and early seedling growth. Plants containing AtCYN or OsCYN exhibited resistance to KCNO stress, which demonstrated that one role of cyanases in plants is detoxification. Transcription level of AtCYN was higher in the flower than in other organs of Arabidopsis. AtCYN transcription was not significantly affected by KCNO treatment in Arabidopsis, but was induced by salt stress. This research broadens our knowledge on plant detoxification of cyanate via cyanase.
authors.affiliation: State Key Laboratory of Protein and Plant Gene Research, Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology, College of Life Sciences, Peking University, Beijing, People's Republic of China.
authors.forename: Dan
authors.initials: D
authors.lastname: Qian
authors.forename: Lin
authors.initials: L
authors.lastname: Jiang
authors.forename: Lu
authors.initials: L
authors.lastname: Lu
authors.forename: Chunhong
authors.initials: C
authors.lastname: Wei
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology*; D019759:Carbon-Nitrogen Lyases / Q000737:chemistry / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology*; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013696:Temperature
chemical_list: D010940:Plant Proteins; C019953:cyanate hydrolase; D019759:Carbon-Nitrogen Lyases