Harafuji 1980 J Biochem
Harafuji H, Ogawa Y (1980) Re-examination of the apparent binding constant of ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid with calcium around neutral pH. J Biochem 87(5):1305-12. |
Harafuji H, Ogawa Y (1980) J Biochem
Abstract: 1. The apparent binding constant (Kapp(Ca-G)) for GEDTA (ethylene glycol bis(Ξ²-aminoethyl ether)-N, N, N', N'-tetraacetic acid, EGTA) to calcium was determined under conditions of biological significance in the presence of various kinds of pH-buffering agents, using murexide or tetramethylmurexide as a Ca indicator. 2. The value of Kapp(Ca-G) at pH 6.80 was 1.0Γ106M-1 at an ionic strength of 0.114 at 20Β°C, irrespective of the type of pH-buffering ions. This value is similar to that of Allen, Blinks and Prendergast (1977) (Science 196, 996-998), but still half that calculated from the results of Schwarzenbach, Senn and Anderegg (1957) (Helv. Chim. Acta 40, 1886-1900). 3. The value of Kapp(Ca-G) varied according to the following equation as the ionic strength (I) was varied from 0.039 to 0.264: log Kapp(Ca-G)=6.460-[2_??_I/(1+_??_I)-0.4ΓI] (pH 6.80, 20Β°C) 4. The discrepancy between the present results and previous ones (Ogawa, Y. (1968) J. Biochem. 64, 255-257) may have been due to inadequate regulation of the temperature of the reaction medium in the previous determinations, during which an increase in the temperature of the solution may have occurred. An increase of temperature causes a decrease in the pH of the solution in the presence of histidine, imidazole or Tris-maleate, but causes very little change of pH in the presence of phosphate or maleate. 5. The association rate constant for GEDTA with calcium was determined by the stoppedflow method in solutions containing 100mM KCl and 20mM pH-buffering ions at 20Β°C: the values obtained were 1.4Γ106M-1s-1 in the presence of MOPS-KOH at pH 6.80; 3.0Γ106M-1s-1 with imidazole at pH 6.80; 1.0Γ106M-1s-1 with Tris-maleate at pH 6.80. β’ Keywords: calcium, EGTA, aparent biding constant β’ Bioblast editor: Cecatto C
Labels: MiParea: Instruments;methods
Regulation: Calcium