Biotin-dependent expression of the asialoglycoprotein receptor in HepG2

J. C. Collins, Elisabeth M. Paietta, R. Green, A. G. Morell, R. J. Stockert

Research output: Contribution to journalArticle

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Abstract

The asialoglycoprotein receptor (AsGR) is characteristic of fully differentiated hepatocytes. AsGR expression in confluent cultures of HepG2 cells grown in minimal essential medium (MEM) requires a 300-350-dalton dialyzable fraction of fetal bovine serum (FBS). Addition to dialyzed FBS (dFBS) of 10-7 M biotin or biocytin (M(r) 372) permitted full expression of AsGR by HepG2. Affinity chromatography of FBS on streptavidin-Sepharose abolished its ability to support AsGR production. The bound material, when released by heat denaturation and resolved by thin layer chromatography, yielded three cinnamaldehyde-positive components, of which the major detectable one migrates with authentic biocytin and reconstitutes dFBS. Sera from several species, which do not support AsGR production by HepG2, contain less than 10% biotin found in FBS as determined by direct enzyme-linked immunosorbent assay. These results indicate that biotin or a derivative is the low molecular weight serum factor of FBS required for expression of AsGR. Isolation of messenger RNA from HepG2 revealed no difference in AsGR transcripts when cells were grown in MEM-10% FBS or MEM-10% dFBS. Thus a biotin-dependent post-transcriptional event permits the ultimate expression of the AsGR by HepG2 cells.

Original languageEnglish (US)
Pages (from-to)11280-11283
Number of pages4
JournalJournal of Biological Chemistry
Volume263
Issue number23
StatePublished - 1988

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Asialoglycoprotein Receptor
Biotin
Serum
Hep G2 Cells
Affinity chromatography
Thin layer chromatography
Immunosorbents
Denaturation
Thin Layer Chromatography
Affinity Chromatography
Cell culture
Hepatocytes
Assays
Hot Temperature
Molecular Weight
Enzyme-Linked Immunosorbent Assay
Molecular weight
Cells
Derivatives
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Collins, J. C., Paietta, E. M., Green, R., Morell, A. G., & Stockert, R. J. (1988). Biotin-dependent expression of the asialoglycoprotein receptor in HepG2. Journal of Biological Chemistry, 263(23), 11280-11283.

Biotin-dependent expression of the asialoglycoprotein receptor in HepG2. / Collins, J. C.; Paietta, Elisabeth M.; Green, R.; Morell, A. G.; Stockert, R. J.

In: Journal of Biological Chemistry, Vol. 263, No. 23, 1988, p. 11280-11283.

Research output: Contribution to journalArticle

Collins, JC, Paietta, EM, Green, R, Morell, AG & Stockert, RJ 1988, 'Biotin-dependent expression of the asialoglycoprotein receptor in HepG2', Journal of Biological Chemistry, vol. 263, no. 23, pp. 11280-11283.
Collins, J. C. ; Paietta, Elisabeth M. ; Green, R. ; Morell, A. G. ; Stockert, R. J. / Biotin-dependent expression of the asialoglycoprotein receptor in HepG2. In: Journal of Biological Chemistry. 1988 ; Vol. 263, No. 23. pp. 11280-11283.
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