Abstract
Fluorimetric or spectrophotometric titrations with the appropriate cations gave Kd values of 2.9 +/- 0.2 nM and 89 +/- 5 microM respectively for the Ca2+ and Mg2+ complexes of quin 2 at pH 7.5. Mixtures of quin 2 and vitamin D-dependent Ca2+-binding protein from pig duodenum were titrated fluorimetrically with Ca2+ in the absence or presence of Mg2+. These measurements were used with the Kd values of the Ca2+ and Mg2+ complexes of quin 2 to obtain Kd or apparent Kd values for Ca2+-protein complexes ranging from 5 nM to 5 microM with good accuracy.
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Selected References
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- Barker W. C., Dayhoff M. O. Evolution of homologous physiological mechanisms based on protein sequence data. Comp Biochem Physiol B. 1979;62(1):1–5. doi: 10.1016/0305-0491(79)90002-6. [DOI] [PubMed] [Google Scholar]
- Bryant D. T., Andrews P. A simple procedure for purifying mammalian duodenal Ca2+-binding proteins on a 100 mg scale and an investigation of the stoichiometry of their high-affinity binding of Ca2+ ions. Biochem J. 1983 Jun 1;211(3):709–716. doi: 10.1042/bj2110709. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bryant D. T., Andrews P. Investigation of the binding of Ca2+, Mg2+, Mn2+ and K+ to the vitamin D-dependent Ca2+-binding protein from pig duodenum. Biochem J. 1984 Apr 1;219(1):287–292. doi: 10.1042/bj2190287. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hesketh T. R., Smith G. A., Moore J. P., Taylor M. V., Metcalfe J. C. Free cytoplasmic calcium concentration and the mitogenic stimulation of lymphocytes. J Biol Chem. 1983 Apr 25;258(8):4876–4882. [PubMed] [Google Scholar]
- Hofmann T., Kawakami M., Hitchman A. J., Harrison J. E., Dorrington K. J. The amino acid sequence of porcine intestinal calcium-binding protein. Can J Biochem. 1979 Jun;57(6):737–748. doi: 10.1139/o79-092. [DOI] [PubMed] [Google Scholar]
- Kim Y. S., Padilla G. M. Determination of free Ca ion concentrations with an ion-selective electrode in the presence of chelating agents in comparison with calculated values. Anal Biochem. 1978 Sep;89(2):521–528. doi: 10.1016/0003-2697(78)90381-0. [DOI] [PubMed] [Google Scholar]
- Murakami K., Andree P. J., Berliner L. J. Metal ion binding to alpha-lactalbumin species. Biochemistry. 1982 Oct 26;21(22):5488–5494. doi: 10.1021/bi00265a017. [DOI] [PubMed] [Google Scholar]
- Permyakov E. A., Yarmolenko V. V., Kalinichenko L. P., Morozova L. A., Burstein E. A. Calcium binding to alpha-lactalbumin: structural rearrangement and association constant evaluation by means of intrinsic protein fluorescence changes. Biochem Biophys Res Commun. 1981 May 15;100(1):191–197. doi: 10.1016/s0006-291x(81)80081-2. [DOI] [PubMed] [Google Scholar]
- Tsien R. Y. New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures. Biochemistry. 1980 May 27;19(11):2396–2404. doi: 10.1021/bi00552a018. [DOI] [PubMed] [Google Scholar]
- Tsien R. Y., Pozzan T., Rink T. J. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol. 1982 Aug;94(2):325–334. doi: 10.1083/jcb.94.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
