Characterization of the basement membrane glycoprotein entactin synthesized in a baculovirus expression system

T. Tsao, J. C. Hsieh, M. E. Durkin, C. Wu, S. Chakravarti, L. J. Dong, M. Lewis, A. E. Chung

Research output: Contribution to journalArticle

Abstract

A cDNA clone encoding the entire open reading frame of mouse entactin was constructed. This cDNA was inserted into Autographa californica multiple nuclear polyhedrosis virus and expressed in Spodoptera frugiperda, Sf9, insect cells. The recombinant entactin was produced in large quantities under the control of the polyhedrin promoter. Amino terminus sequence analysis indicated that the signal peptide in the pre-protein was correctly cleaved. Polyclonal antibodies prepared against either the naturally occurring or recombinant entactin cross-reacted with both molecules in the native and denatured states. The recombinant molecule promoted cell attachment and exhibited a calcium- and temperature-dependent self-aggregation. These properties reflect those of the naturally occurring mouse molecule. Although entactin is an extracellular matrix component that is normally exocytosed, the vast majority of the molecules were localized inside the cell by immunoelectron microscopy where the molecules formed insoluble aggregates.

Original languageEnglish (US)
Pages (from-to)5188-5191
Number of pages4
JournalJournal of Biological Chemistry
Volume265
Issue number9
StatePublished - 1990
Externally publishedYes

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Baculoviridae
Membrane Glycoproteins
Basement Membrane
Molecules
Complementary DNA
Nucleopolyhedrovirus
Sf9 Cells
Spodoptera
Immunoelectron Microscopy
Protein Sorting Signals
Open Reading Frames
Extracellular Matrix
Sequence Analysis
Insects
Viruses
Clone Cells
Calcium
Microscopic examination
Agglomeration
Temperature

ASJC Scopus subject areas

  • Biochemistry

Cite this

Tsao, T., Hsieh, J. C., Durkin, M. E., Wu, C., Chakravarti, S., Dong, L. J., ... Chung, A. E. (1990). Characterization of the basement membrane glycoprotein entactin synthesized in a baculovirus expression system. Journal of Biological Chemistry, 265(9), 5188-5191.

Characterization of the basement membrane glycoprotein entactin synthesized in a baculovirus expression system. / Tsao, T.; Hsieh, J. C.; Durkin, M. E.; Wu, C.; Chakravarti, S.; Dong, L. J.; Lewis, M.; Chung, A. E.

In: Journal of Biological Chemistry, Vol. 265, No. 9, 1990, p. 5188-5191.

Research output: Contribution to journalArticle

Tsao, T, Hsieh, JC, Durkin, ME, Wu, C, Chakravarti, S, Dong, LJ, Lewis, M & Chung, AE 1990, 'Characterization of the basement membrane glycoprotein entactin synthesized in a baculovirus expression system', Journal of Biological Chemistry, vol. 265, no. 9, pp. 5188-5191.
Tsao, T. ; Hsieh, J. C. ; Durkin, M. E. ; Wu, C. ; Chakravarti, S. ; Dong, L. J. ; Lewis, M. ; Chung, A. E. / Characterization of the basement membrane glycoprotein entactin synthesized in a baculovirus expression system. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 9. pp. 5188-5191.
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