| Subcellular localization and partial purification of the 25-hydroxyvitamin D3 1-hydroxylation reaction in the chick myelomonocytic cell line HD-11. | |
| | |
MedLine Citation:
|
PMID: 8384398 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Hypercalcemia in human granuloma-forming diseases like sarcoidosis results from the endogenous overproduction of 1,25-dihydroxyvitamin D [1,25-(OH)2D] by disease-activated tissue macrophages. The recent identification of an immortalized chick myelomonocytic cell line, HD-11, that constitutively expresses a 25-hydroxyvitamin D (25-OHD) 1-hydroxylation reaction has alleviated dependence on studying primary macrophage cultures with no replicative potential in vitro. In these experiments we established conditions for the maximal expression of the HD-11 cell 25-OHD3-1-hydroxylation reaction and localized this activity to the mitochondrial fraction. On a per cell basis, the activity of HD-11 cell 25-OHD3 1-hydroxylation reaction was comparable to that in primary cultures of chick renal tubular epithelial cells, which express the authentic renal 25-OHD3 1-hydroxylase. Maximal product yield was achieved after incubation of HD-11 cells with 200 nM 25-OHD3 for 3 h. Although adherent monolayers possessed 3- to 4-fold more capacity for hormone production than cells in suspension, suspended cells exhibited easily detectable 25-OHD3 catalytic activity (0.58 +/- 0.08 pmol per 10(6) cells per h; +/- SEM), 50% of which remained solubilized in a sonicate of suspended cells cleared of nuclei and plasma membrane. Subcellular localization disclosed 91% of the residual activity to be concentrated in the mitochondrial subfraction. A detergent-solubilized extract of this mitochondrial subfraction contained 1.9 +/- 0.3 pmol 1,25-(OH)2D3 synthetic capacity per mg protein. The catalytic activity (1-hydroxylase activity) was concentrated 20.2-fold after chromatography on octyl-amino agarose and was associated with 0.054 nmol cytochrome P450 per mg protein.(ABSTRACT TRUNCATED AT 250 WORDS) |
| | |
Authors:
|
S Shany; S Y Ren; J E Arbelle; T L Clemens; J S Adams |
Related Documents
:
|
6941298 - Increased mitochondrial uptake of rhodamine 123 during lymphocyte stimulation. 16973208 - Mitochondrial damage and intralysosomal degradation in cellular aging. 16127068 - Oxazolidinones inhibit cellular proliferation via inhibition of mitochondrial protein s... 15965098 - Enhancement of cisplatin-induced apoptosis and caspase 3 activation by depletion of mit... 1127378 - Identification of a novel cell type in peripheral lymphoid organs of mice. iv. identifi... 6714348 - Actions of cholinergic drugs on cells in the interpeduncular nucleus. |
Publication Detail:
|
Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
|
Title: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research Volume: 8 ISSN: 0884-0431 ISO Abbreviation: J. Bone Miner. Res. Publication Date: 1993 Mar |
Date Detail:
|
Created Date: 1993-04-21 Completed Date: 1993-04-21 Revised Date: 2007-11-14 |
Medline Journal Info:
|
Nlm Unique ID: 8610640 Medline TA: J Bone Miner Res Country: UNITED STATES |
Other Details:
|
Languages: eng Pagination: 269-76 Citation Subset: IM |
Affiliation:
|
Department of Medicine, Cedars-Sinai Research Institute, University of California Los Angeles School of Medicine. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Animals Avian leukosis virus Calcitriol / isolation & purification*, metabolism Chickens Chromatography, Affinity Cytochrome P-450 Enzyme System / chemistry Hydroxylation Kidney Tubules / cytology, metabolism Macrophages / metabolism* Tumor Cells, Cultured |
| Grant Support | |
ID/Acronym/Agency:
|
DK-33139/DK/NIDDK NIH HHS |
| Chemical | |
Reg. No./Substance:
|
32222-06-3/Calcitriol; 9035-51-2/Cytochrome P-450 Enzyme System |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: An intradermal test for the diagnosis of BHV-1 infection. The effect of repeated testing on the immu...
Next Document: Cross talk of dual-signal transduction systems in the regulation of DNA synthesis by parathyroid hor...