HEME BINDING TO HEP-G2 HUMAN HEPATOMA-CELLS

被引:16
作者
GALBRAITH, RA
机构
[1] Department of Metabolism/Pharmacology, The Rockefeller University Hospital, New York, NY
关键词
D O I
10.1016/0168-8278(90)90137-G
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Utilizing high specific activity [55Fe]hemin, the interaction of heme with monolayer cultures of human Hep G2 hepatoblastoma cells was examined. Initial characterization was performed at 4 °C to minimize the possibility of heme internalization. Specific binding of [55Fe]hemin at 4 °C reached equilibrium within 6 h and was 66% dissociable after 18 h in the presence of fresh binding buffer. Stereospecificity of the binding interaction was evidenced by variable degrees of competition with different metalloporphyrins. Scatchard analysis revealed a Ka of 0.01 nM-1 and the number of binding sites per cell was approx. 200 000. Elevation of the temperature during binding to 22 °C resulted in a small decrease in the affinity of the interaction and the apparent number of sites per cell was doubled, possibly due to artefactual elevation of the cell-associated radioactivity secondary to intracellular accumulation. These data are interpreted to indicate the presence of a specific heme receptor on the plasmalemma of Hep G2 cells. © 1990.
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页码:305 / 310
页数:6
相关论文
共 31 条
[1]   SPECTROSCOPIC STUDY OF HEMIN - HUMAN SERUM-ALBUMIN SYSTEM [J].
BEAVEN, GH ;
CHEN, SH ;
DALBIS, A ;
GRATZER, WB .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1974, 41 (03) :539-546
[2]   INDUCTION OF HEME-SYNTHESIS IN HEP G2 HUMAN HEPATOMA-CELLS BY DIMETHYLSULFOXIDE - A TRANSCRIPTIONALLY ACTIVATED EVENT [J].
GALBRAITH, RA ;
SASSA, S ;
KAPPAS, A .
BIOCHEMICAL JOURNAL, 1986, 237 (02) :597-600
[3]   REGULATION OF HEME-SYNTHESIS IN HEPG2 HUMAN HEPATOMA-CELLS BY DIMETHYL-SULFOXIDE [J].
GALBRAITH, RA ;
SASSA, S ;
FUJITA, H .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1988, 153 (02) :869-874
[4]   HEME BINDING TO LEISHMANIA-MEXICANA-AMAZONENSIS [J].
GALBRAITH, RA ;
MCELRATH, MJ .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1988, 29 (01) :47-53
[5]  
GALBRAITH RA, 1985, J BIOL CHEM, V260, P2198
[6]  
GRASBECK R, 1979, SCAND J HAEMATOL, V23, P5
[7]   SPECTRAL AND OTHER STUDIES ON THE INTESTINAL HEME RECEPTOR OF THE PIG [J].
GRASBECK, R ;
MAJURI, R ;
KOUVONEN, I ;
TENHUNEN, R .
BIOCHIMICA ET BIOPHYSICA ACTA, 1982, 700 (02) :137-142
[8]  
HERSHKO C, 1972, J LAB CLIN MED, V80, P624
[9]   PORPHYRIN BINDING BY THE SURFACE ARRAY VIRULENCE PROTEIN OF AEROMONAS-SALMONICIDA [J].
KAY, WW ;
PHIPPS, BM ;
ISHIGURO, EE ;
TRUST, TJ .
JOURNAL OF BACTERIOLOGY, 1985, 164 (03) :1332-1336
[10]   HUMAN HEPATOCELLULAR-CARCINOMA CELL-LINES SECRETE THE MAJOR PLASMA-PROTEINS AND HEPATITIS-B SURFACE-ANTIGEN [J].
KNOWLES, BB ;
HOWE, CC ;
ADEN, DP .
SCIENCE, 1980, 209 (4455) :497-499