title: Ribonuclease inhibitors in Malus x domestica (common apple): isolation and partial characterization.
abstract: A ribonuclease inhibitory activity was detected in the fruits of common apple, Malus x domestica, cv. Fuji, and purified by affinity chromatography on ribonuclease A-Sepharose. It inhibited hydrolysis of cyclic-2':3'-CMP by bovine pancreatic ribonuclease A with an apparent inhibition constant of about 5 x 10(-8) M. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of the purified protein gave two peaks corresponding to the mass numbers of 55,658 and 62,839, while three bands of 43-, 34-, and 21-kDa were detected by SDS-PAGE. These results suggested that the inhibitor preparation was a mixture of two proteins comprised of 43- and 21-kDa subunits or of 34- and 21-kDa subunits. Attempts to separate these two proteins were unsuccessful. Amino acid composition and N-terminal amino acid sequence of these subunits were also identified and N-terminal sequences showed some similarity to that of cottonseed storage globulin. The significance of the presence of ribonuclease inhibitors in apple fruits is not clear, but it might allow some speculation about their possible involvement in the control of the self-incompatibility ribonuclease of Rosaceae plants.
authors.affiliation: Department of Biology, Faculty of Science, Niigata University, Ikarashi, Niigata 950-2181, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Kosuge
authors.forename: Mamoru
authors.initials: M
authors.lastname: Isemura
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Sumiko
authors.initials: S
authors.lastname: Odani
authors.forename: Shoji
authors.initials: S
authors.lastname: Odani
mesh_terms: D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D004791:Enzyme Inhibitors / Q000302:isolation & purification*; D027845:Malus / Q000201:enzymology*; D008970:Molecular Weight; D012260:Ribonucleases / Q000037:antagonists & inhibitors*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D004791:Enzyme Inhibitors; D012260:Ribonucleases