Denitration of peroxynitrite-treated proteins by "protein nitratases" from dog prostate.
Putative "protein nitratases," which catalyze denitration of peroxynitrite (PN)-treated, proteins, were detected in the crude extract of dog prostate. Nitratase activity was monitored by the decreased intensity of nitrotyrosine immunoreactive-bands in Western blot and increased nitrate level in dialysate of incubation mixture, which contained prostate crude extract, protease inhibitors and a PN-treated substrate, such as treated histone (III-S), BSA, invertase, or polylysine. Nitratases were activated by preincubation with m-calpain/Ca2+. Furthermore, after denitration, the activity of PN/DTT-treated invertase decreased to the similar activity level of DTT-treated invertase. At least two different types of nitratases may occur: type I, reductant-dependent, and type II, reductant-independent.
Division of Science and Mathematics, Bethune-Cookman College, Daytona Beach, Florida 32114, USA.
W N
WN
Kuo
R N
RN
Kanadia
V P
VP
Shanbhag
D000818:Animals; D015153:Blotting, Western; D004229:Dithiothreitol; D004285:Dogs; D004789:Enzyme Activation; D005982:Glutathione Transferase / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry; D006657:Histones / Q000737:chemistry; D008297:Male; D009566:Nitrates; D010088:Oxidoreductases / Q000378:metabolism*; D011107:Polylysine / Q000737:chemistry; D011467:Prostate / Q000201:enzymology*; D012709:Serum Albumin / Q000737:chemistry; D014020:Tissue Extracts / Q000737:chemistry; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis; D043324:beta-Fructofuranosidase
D006657:Histones; D009566:Nitrates; D012709:Serum Albumin; D014020:Tissue Extracts; D011107:Polylysine; C039642:peroxynitric acid; C002744:3-nitrotyrosine; D014443:Tyrosine; D010088:Oxidoreductases; C045258:nitratase; D005982:Glutathione Transferase; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D004229:Dithiothreitol