| Intracellular calcium measurements by 19F NMR of fluorine-labeled chelators. | |
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MedLine Citation:
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PMID: 6417665 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Symmetrically substituted difluoro derivatives of 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (nFBAPTA) show large 19F NMR chemical shifts on chelating divalent cations. The complexes of Ca2+ with 4FBAPTA and 5FBAPTA show fast and slow exchange behavior, respectively, and the chemical shift or the areas of the resonances from the free and complexed forms can be used to determine the free Ca2+ concentration. The measurement of the free Ca2+ concentration by either ligand is unaffected by free Mg2+ concentrations less than 10 mM, by pH 6-8, or by contaminating divalent ions of high affinity (Zn2+, Fe2+, Mn2+). The tetraacetoxymethyl ester derivative of 5FBAPTA was used to load mouse thymocytes with 5FBAPTA to intracellular concentrations of 1 mM, and the 19F spectrum indicated a free intracellular Ca2+ concentration [( Ca]i) of 250 nM. The [Ca]i was increased to 350 nM by addition of succinylated concanavalin A at mitogenic concentrations, and the addition of A23187 saturated the intracellular chelator with Ca2+ from the external medium. The method provides a measurement of [Ca]i and other divalent cation concentrations with direct identification of the ionic species chelated. |
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Authors:
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G A Smith; R T Hesketh; J C Metcalfe; J Feeney; P G Morris |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 80 ISSN: 0027-8424 ISO Abbreviation: Proc. Natl. Acad. Sci. U.S.A. Publication Date: 1983 Dec |
Date Detail:
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Created Date: 1984-01-27 Completed Date: 1984-01-27 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 7505876 Medline TA: Proc Natl Acad Sci U S A Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 7178-82 Citation Subset: IM |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Calcium / analysis* Chelating Agents* Egtazic Acid* / analogs & derivatives Ethylene Glycols* Magnesium / analysis Magnetic Resonance Spectroscopy / methods Mice Mice, Inbred BALB C T-Lymphocytes / analysis* Thymus Gland / analysis |
| Chemical | |
Reg. No./Substance:
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0/Chelating Agents; 0/Ethylene Glycols; 67-42-5/Egtazic Acid; 7439-95-4/Magnesium; 7440-70-2/Calcium; 85233-21-2/5,5'-difluoro-1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; 85233-22-3/4,4'-difluoro-1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid |
| Comments/Corrections | |
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