The expression of milligram amounts of functional human 1,25-dihydroxyvitamin D3 receptor in a bacterial expression system

Rajiv Kumar, Janet Schaefer, Eric Wieben

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

We expressed milligram amounts of functional human 1,25-dihydroxyvitamin D3 receptor in a bacterial expression system in which the cloned cDNA for the hVDR was expressed under the control of bacterial T7 polymerase. The hVDR protein comprised approximately 60% of total bacterial protein. It migrated on polyacrylamide-sodium dodecyl sulfate gels with an Mr of 48,000. It had the predicted amino acid composition and amino acid sequence analysis. The expressed protein was bound by 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) with a Kd in the nanomolar range. It sedimented on sucrose density gradients at 3.5S. Furthermore, the expressed protein bound to the osteocalcin vitamin D response element (VDRE) as assessed by a gel mobility shift assay. The expression of large amounts of hVDR protein should allow for the use of this protein in structure-function and x-ray crystallography studies.

Original languageEnglish (US)
Pages (from-to)1417-1423
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume189
Issue number3
DOIs
StatePublished - Dec 30 1992

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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