title: Purification and characterization of two membrane bound serine proteinases from rat liver microsomes active in degradation of cytochrome P450.
abstract: Two serine proteinases capable of digesting cytochrome P4502E1 (CYP2E1) have been purified from sodium cholate solubilized rat liver microsomal membranes. After chromatography on hydroxyapatite, DEAE-Sepharose chromatography resolved the CYP2E1-degrading activity into two peaks, and the two proteinases were finally purified on benzamidine-Sepharose. Both have a M(r) of 32,000 on SDS-PAGE, are optimally active at pH 8, and show a susceptibility to inhibitors typical of serine proteinases. CYP2E1 degradation patterns exhibited by the proteinases are identical to each other and similar to that observed during the proteolysis of endogenous CYP2E1 in the microsomal membranes, which indicates that the proteinases can degrade CYP2E1 in its native environment. We suggest a role of these proteinases in the rapid phase of cytochrome P450 degradation in the endoplasmic reticulum.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Zhukov
authors.forename: V
authors.initials: V
authors.lastname: Werlinder
authors.forename: M
authors.initials: M
authors.lastname: Ingelman-Sundberg
mesh_terms: D000818:Animals; D019392:Cytochrome P-450 CYP2E1; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012697:Serine Endopeptidases / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D011480:Protease Inhibitors; D003577:Cytochrome P-450 Enzyme System; D019392:Cytochrome P-450 CYP2E1; D010089:Oxidoreductases, N-Demethylating; D012697:Serine Endopeptidases