Identification and Partial Purification of Ethanol-Induced Hemoproteins in Human Liver

사람의 간에서 Ethanol에 의해 유발되는 hemoprotein들의 확인 및 부분정제

  • Park, Sung-Woo (Division of Chemical Analysis, The National Institute of Scientific Investigation) ;
  • Seo, Bae-Seok (Department of Chemistry, Catholic University Medical College) ;
  • Jin, Kwang-Ho (Division of Chemical Analysis, The National Institute of Scientific Investigation)
  • 박성우 (국립과학수사연구소 화학분석과) ;
  • 서배석 (가톨릭의과대학 화학과) ;
  • 진광호 (국립과학수사연구소 화학분석과)
  • Received : 1994.11.25
  • Published : 1995.06.25

Abstract

To Purify hemoproteins showing from 218nm absorbance, crude liver extract of human with hepatocirrhosis was treated with Triton N-101. Hemoproteins were purified by modification of Mohamed's method. This crude extract was applied to Octyl-Sepharose CL-4B column and the step elution was performed with 0.06% Lubrol PX and 0.25% Lubrol PX. The absorption of effluents were examined at 418nm and two peaks were appeared(Fig. 2). Hemoproteins were purified from Hyydroxyapatite and DEAE-Sephadex A-25 columns which the first peak was applied to(Fig. 3, 4). In death with suddenly, purified hemoproteins with 62 and 45kDa were obtained from 12.5% SDS-PAGE. In death with hepatocirrhosis, purified hemoprotein with 54kDa was obtainded from 12.5% SDS-PAGE(Fig. 5). Cytochrome P450 was purified to a specific content of 20.8nmol/mg protein with a recovery of about 4.1%. Absorbance maximum of these hemoproteins were 446nm at UV spectruum(Fig. 6).

Keywords

Death with hepatocirrhosis;Hemoproteins;Column;SDS-PAGE;Cytochrome P450