Presence of oxidized protein hydrolase in human cell lines, rat tissues, and human/rat plasma

Citation
T. Fujino et al., Presence of oxidized protein hydrolase in human cell lines, rat tissues, and human/rat plasma, J BIOCHEM, 127(2), 2000, pp. 307-313
Citations number
42
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
JOURNAL OF BIOCHEMISTRY
ISSN journal
0021-924X → ACNP
Volume
127
Issue
2
Year of publication
2000
Pages
307 - 313
Database
ISI
SICI code
0021-924X(200002)127:2<307:POOPHI>2.0.ZU;2-B
Abstract
Oxidized protein hydrolase (OPH), an 80 kDa serine protease whose activity is inhibited by diisopropyl fluorophosphate (DFP), has been isolated from h uman erythrocytes [Fujino, T. ct at (1998) J. Biochem. 124, 1077-1085]. The presence of OPH in various biological samples was examined by enzyme-linke d immunosorbent assay (ELISA) and immunoblotting using an anti-OPH antibody raised against OPH purified from human erythrocytes, and by [H-3]DFP-label ing and successive SDS-PAGE/fluorography. Solubilized samples of human cell lines including K-562 cells, THP-1 cells and Jurkat cells, and rat tissues including brain, heart, liver, kidney, and testis, inhibited the anti-OPH antibody binding to OPH in ELISA. Immunoblotting of lysates of K-562 cells, THP-1 cells and Jurkat cells showed four immunoreactive protein bands incl uding an 80 kDa protein. Immunoprecipitation of the [H-3]DFP-labeled K-562 cell lysate and successive SDS-PAGE/fluorography showed the presence of onl y the 80 kDa DFP-reactive protein with OPH antigenic activity. The level of the 80 kDa immunoreactive protein in K-562 cells rose as the cells differe ntiated toward erythrocytes. Immunoblotting of human and rat plasma showed two immunoreactive protein bands, including the 80 kDa protein, and SDSPAGr E/fluorography of [H-3]DFP-labeled rat and human plasma showed the presence of only the 80 kDa DFP-reactive protein, The results indicate that OPH is present in a wide variety of biological samples.