The RNA hydrolysis and the cytokinin binding activities of PR-10 proteins are differently performed by two isoforms of the Pru p 1 peach major allergen and are possibly functionally related.
PR-10 proteins are a family of pathogenesis-related (PR) allergenic proteins playing multifunctional roles. The peach (Prunus persica) major allergen, Pru p 1.01, and its isoform, Pru p 1.06D, were found highly expressed in the fruit skin at the pit hardening stage, when fruits transiently lose their susceptibility to the fungal pathogen Monilinia spp. To investigate the possible role of the two Pru p 1 isoforms in plant defense, the recombinant proteins were expressed in Escherichia coli and purified. Light scattering experiments and circular dichroism spectroscopy showed that both proteins are monomers in solution with secondary structures typical of PR-10 proteins. Even though the proteins do not display direct antimicrobial activity, they both act as RNases, a function possibly related to defense. The RNase activity is different for the two proteins, and only that of Pru p 1.01 is affected in the presence of the cytokinin zeatin, suggesting a physiological correlation between Pru p 1.01 ligand binding and enzymatic activity. The binding of zeatin to Pru p 1.01 was evaluated using isothermal titration calorimetry, which provided information on the stoichiometry and on the thermodynamic parameters of the interaction. The structural architecture of Pru p 1.01 and Pru p 1.06D was obtained by homology modeling, and the differences in the binding pockets, possibly accounting for the observed difference in binding activity, were evaluated.
Zubini
Paola
P
Department of Agri-Food Protection and Improvement, CRIOF , University of Bologna, 40127 Bologna, Italy.
Zambelli
Barbara
B
Musiani
Francesco
F
Ciurli
Stefano
S
Bertolini
Paolo
P
Baraldi
Elena
E
D000485:Allergens / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D003583:Cytokinins / Q000378:metabolism*; D006868:Hydrolysis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D027861:Prunus / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D018749:RNA, Plant / Q000378:metabolism*; D016415:Sequence Alignment; D015026:Zeatin / Q000378:metabolism
D000485:Allergens; D052179:Antigens, Plant; D003583:Cytokinins; D010940:Plant Proteins; D020033:Protein Isoforms; C403252:Pru p 1 allergen; D018749:RNA, Plant; D015026:Zeatin