Purification and characterization of a nuclease from Lentinus edodes.
An endonuclease with 3'-nucleotidase activity (nuclease Le1) was purified from fruit bodies of Lentinus edodes in a single band on sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The apparent molecular weight of nuclease Le1 was about 27000. The nuclease was inactivated in the presence of ethylenediaminetetraacetic acid (EDTA) and reactivated by the addition of Zn2+. Hydrolysis of poly U by the nuclease showed many intermediate size oligomers prior to the formation of 5'-uridine monophosphate (UMP). Therefore, it was concluded that nuclease Le1 was a Zn(2+)-endonuclease similar to P1-nuclease from Penicillium citrinum. The nuclease was very sensitive to ionic strength, but pH-profiles of the hydrolysis of four 3'-nucleotides were very similar to those of P1 nuclease from P. citrinum.
Shimada
H
H
Department of Microbiology, College of Pharmacy, Nihon University, Chiba, Japan.
Inokuchi
N
N
Koyama
T
T
Irie
M
M
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D002413:Cations, Divalent; D002848:Chromatography, DEAE-Cellulose; D004247:DNA / Q000378:metabolism; D004492:Edetic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D004722:Endoribonucleases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011072:Poly U / Q000378:metabolism; D011124:Polyporaceae / Q000201:enzymology*; D012313:RNA / Q000378:metabolism
D000596:Amino Acids; D002413:Cations, Divalent; D007527:Isoenzymes; D011072:Poly U; D012313:RNA; D004247:DNA; D004492:Edetic Acid; D004722:Endoribonucleases; C022543:ribonuclease T(2)