Impairment of the asialoglycoprotein receptor by ethanol oxidation

被引:13
作者
Clemens, DL [1 ]
Halgard, CM [1 ]
Cole, JR [1 ]
Miles, RM [1 ]
Sorrell, MF [1 ]
Tuma, DJ [1 ]
机构
[1] UNIV NEBRASKA,MED CTR,DEPT INTERNAL MED,OMAHA,NE 68105
关键词
ethanol oxidation; receptor-mediated endocytosis; alcohol dehydrogenase;
D O I
10.1016/S0006-2952(96)00524-2
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
It is well established that ethanol exposure impairs the process of receptor-mediated endocytosis in hepatic cells, although the molecular mechanism(s) and the physiological consequence(s) of this impairment are unclear. Because addressing these mechanistic questions is difficult in vivo, we have developed a recombinant cell line of hepatic origin capable of metabolizing ethanol. In this study, we have used these recombinant cells, designated HAD cells, to investigate the ethanol-induced impairment to the receptor-mediated endocytosis of the hepatic asialoglycoprotein receptor. Comparing the binding of the ligand asialoorosomucoid in both the parental Hep G2 cells and the recombinant HAD cells, maintained in the presence and absence of ethanol, revealed decreased ligand binding in the HAD cells. This impairment was accentuated by prolonging the ethanol exposure, reaching approximately 40% in both surface and total receptor populations by 7 days. Addition of the alcohol dehydrogenase inhibitor pyrazole to the ethanol-containing medium abolished this impairment, indicating that the decreased binding was a result of the alcohol dehydrogenase-mediated oxidation of ethanol. Furthermore, using antibody specific to the asialoglycoprotein receptor, it was demonstrated that the ethanol-induced impairment in ligand binding was a consequence of decreased ligand binding and not a result of diminished receptor numbers. These results indicated that ethanol oxidation was required for the ethanol-induced impairment in ligand binding, and that the reduced ligand binding was a result of a decrease in the ability of the ligand to bind to the receptor. Copyright (C) 1996 Elsevier Science Inc.
引用
收藏
页码:1499 / 1505
页数:7
相关论文
共 26 条
[1]   CELL CELL AND CELL MATRIX INTERACTIONS DIFFERENTIALLY REGULATE THE EXPRESSION OF HEPATIC AND CYTOSKELETAL GENES IN PRIMARY CULTURES OF RAT HEPATOCYTES [J].
BENZEEV, A ;
ROBINSON, GS ;
BUCHER, NLR ;
FARMER, SR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (07) :2161-2165
[2]  
BURWEN SJ, 1991, INTRACELLULAR TRAFFI, P248
[3]  
CASEY CA, 1987, J BIOL CHEM, V262, P2704
[4]   IMPROVED MICRO-FLUOROMETRIC DNA DETERMINATION IN BIOLOGICAL-MATERIAL USING 33258-HOECHST [J].
CESARONE, CF ;
BOLOGNESI, C ;
SANTI, L .
ANALYTICAL BIOCHEMISTRY, 1979, 100 (01) :188-197
[5]   CHANGES IN LIVER-SPECIFIC COMPARED TO COMMON GENE-TRANSCRIPTION DURING PRIMARY CULTURE OF MOUSE HEPATOCYTES [J].
CLAYTON, DF ;
DARNELL, JE .
MOLECULAR AND CELLULAR BIOLOGY, 1983, 3 (09) :1552-1561
[6]   EXPRESSION OF ALEUTIAN MINK DISEASE PARVOVIRUS CAPSID PROTEINS BY A RECOMBINANT VACCINIA VIRUS - SELF-ASSEMBLY OF CAPSID PROTEINS INTO PARTICLES [J].
CLEMENS, DL ;
WOLFINBARGER, JB ;
MORI, S ;
BERRY, BD ;
HAYES, SF ;
BLOOM, ME .
JOURNAL OF VIROLOGY, 1992, 66 (05) :3077-3085
[7]   ESTABLISHMENT OF A RECOMBINANT HEPATIC CELL-LINE STABLY EXPRESSING ALCOHOL-DEHYDROGENASE [J].
CLEMENS, DL ;
HALGARD, CM ;
MILES, RR ;
SORRELL, MF ;
TUMA, DJ .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1995, 321 (02) :311-318
[8]   EXPRESSION OF ALCOHOL-DEHYDROGENASE IN PRIMARY MONOLAYER-CULTURES OF RAT HEPATOCYTES [J].
CRABB, DW ;
LI, TK .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1985, 128 (01) :12-17
[9]   CHRONIC ETHANOL ADMINISTRATION IMPAIRS RECEPTOR-MEDIATED ENDOCYTOSIS OF EPIDERMAL GROWTH-FACTOR BY RAT HEPATOCYTES [J].
DALKE, DD ;
SORRELL, MF ;
CASEY, CA ;
TUMA, DJ .
HEPATOLOGY, 1990, 12 (05) :1085-1091
[10]   KINASE-ACTIVITY CONTROLS THE SORTING OF THE EPIDERMAL GROWTH-FACTOR RECEPTOR WITHIN THE MULTIVESICULAR BODY [J].
FELDER, S ;
MILLER, K ;
MOEHREN, G ;
ULLRICH, A ;
SCHLESSINGER, J ;
HOPKINS, CR .
CELL, 1990, 61 (04) :623-634