Cytochrome P-450 in liver microsomes of streptozotocin-induced diabetic rats: purification and characterization

Biochim Biophys Acta. 1993 Mar 1;1141(2-3):245-52. doi: 10.1016/0005-2728(93)90049-l.

Abstract

Two diabetes-inducible forms of cytochrome P-450, named P-450ST-1 and -ST-2, were purified from the liver microsomes of streptozotocin-diabetic male rats by sodium cholate solubilization, octylamino-Sepharose 4B chromatography and high-performance liquid chromatography with DEAE-5PW and hydroxyapatite columns. The purified P-450 forms gave a single band each on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular weight of 48,500 for P-450ST-1 or 48,000 for P-450ST-2. The CO-reduced spectral maxima of P-450ST-1 and -ST-2 were at 451 nm. The two cytochromes had the low-spin state of heme in the oxidized form. Both P-450ST-1 and -ST-2 catalyzed the metabolism of aniline, benzphetamine, p-nitroanisole, testosterone and aminopyrine. However, the catalytic activity of P-450ST-2 for these substrates was apparently higher than that of ST-1. Analyses of the NH2-terminal amino-acid sequence and Western immunoblot showed that P-450ST-1 and -ST-2 differed structurally from each other. The catalytic activities, molecular weights, NH2-terminal sequences and/or immunochemical properties of P-450ST-1 and -ST-2 did not agree with those of the other cytochrome P-450 forms purified from diabetic rats previously.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Cytochrome P-450 Enzyme System / biosynthesis
  • Cytochrome P-450 Enzyme System / isolation & purification
  • Cytochrome P-450 Enzyme System / metabolism*
  • Diabetes Mellitus, Experimental / enzymology*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Induction
  • Male
  • Microsomes, Liver / enzymology*
  • Molecular Sequence Data
  • Rats
  • Rats, Wistar
  • Streptozocin

Substances

  • Streptozocin
  • Cytochrome P-450 Enzyme System