scholarly journals The effects of calcium ions and pH on bovine prothrombin fragment 1. Intrinsic fluroescence studies

1979 ◽  
Vol 177 (3) ◽  
pp. 879-886 ◽  
Author(s):  
M E Scott ◽  
K A Koehler ◽  
R G Hiskey

The effects of pH and Ca2+ on the intrinsic fluorescence of bovine prothrombin fragment 1 were investigated to deduce the nature of protein functional groups involved in Ca2+ binding to fragment 1. From pH values of 9 to 3, increasing the H3O+ concentration results in quenching of the fluorescence of fragment 1. Reversible pH-titration curves are obtained which appear to consist of two regions. From pH 4 to pH6.5 a broad titration curve is obtained, whereas from pH6.5 to 9 a more pronounced titration behaviour is evidenced by a group or groups on fragment 1 with an apparent pKa of approx. 7.5. In contrast, the apparent association constant for Ca2+ and fragment 1 shows a sharp pH-dependence in the region between pH7 and 8 with tighter Ca2+ binding at higher pH values. A PKa of approx. 7.5 can be estimated for the group or groups on fragment 1 linked to the tight binding of Ca2+. Both H3O+ and Ca2+ result in blue-shifts in the wave-lengths of fragment-1 emission. These results are interpreted in terms of H+ - and Ca2+ - induced changes in the conformation of fragment 1 as a result of surface-charge neutralization.

2003 ◽  
Vol 807 ◽  
Author(s):  
Y. Zhang ◽  
E. Loi ◽  
M. Blackford ◽  
T. McLeod ◽  
H. Li ◽  
...  

ABSTRACTThe dissolution of the thorium analogue of brannerite (ThTi2O6-I) and U(IV)/U(V) doped Th-brannerite (Th0.97U0.03Ti2O6-II and Th0.955U0.03Ca0.015Ti2O6-III) in aqueous media under atmospheric conditions has been studied to elucidate the effects of pH and uranium valence state on the dissolution rate.The dissolution of I is nearly stoichiometric but slightly preferential release of U occurs for II and preferential release of Ca and U occur for III. The V-shape pH dependence previously observed for U-brannerite only occurs for U (not other matrix elements) for II, indicating that the pH dependence is related to the U oxidation state upon dissolution. The normalised U dissolution rates of III are nearly an order of magnitude higher than those of II for pH values over 3, suggesting brannerite is less durable with U(V) doping. TEM examination of specimens after leaching revealed few surface alteration products, which is consistent with the nearly stoichiometric dissolution of thorium brannerite.


1969 ◽  
Vol 114 (4) ◽  
pp. 863-870 ◽  
Author(s):  
R W Stoddart ◽  
I. P. C. Spires ◽  
K F Tipton

1. The specimen of polygalacturonic acid used in these studies was shown to contain very little neutral sugar, methyl ester groups or ash, and only residues of galacturonic acid. Its electrophoretic homogeneity was examined in pyridine–acetic acid buffer at pH6·5 and in borate buffer at pH9·2. The distribution of effective particle weights was shown to be fairly narrow. 2. The pH-titration curve of the polymer gave a pK value of 3·7. 3. The interaction of the polymer with Ruthenium Red was studied and titration curves were obtained for the spectral shifts associated with the formation of a complex. 4. Optical-rotatory-dispersion studies showed that the Drude constant, λc, was dependent on pH. 5. Polygalacturonic acid was shown to display non-Newtonian properties in solution and to have an anomalously high relative specific viscosity at low concentrations. 6. Studies were made of the pH-dependence of the sedimentation coefficient of the polymer. 7. These results are discussed in terms of the structure of the molecule and their relevance to the properties of pectic substances.


1979 ◽  
Vol 41 (02) ◽  
pp. 296-308
Author(s):  
Daniel J Schaefer ◽  
Mary E Scott ◽  
Don A Gabriel ◽  
Jean L Gerth ◽  
David L Aronson ◽  
...  

SummaryVariation of pH strongly affects the fluorescence intensity of human prothrombin fragment-1 in a manner suggesting contributions from a number of protropic equilibria including groups with apparent pKa values near 3.0. These results suggest a structural role for pKa 1 of γ-carboxyglutamic acid moieties. Added calcium ions (9 mM calcium chloride) quench the fluorescence titration curve.uniformly above pH 4. Below pH 4, however, the titration curve in the presence of calcium ions suggests that calcium-ion-dependent processes leading to fluorescence quenching are pH-dependent. Upon back titration of human fragment-1, from pH 9, hysteresis is observed.Human prothrombin fragment-2 fluorescence titration curves are relatively broad at low pH suggesting the titration of normal carboxyl groups. The titration curves of fragment-2 are not affected by the presence of calcium ions, and hysteresis occurs upon back titration from low pH values. Circular dichroism (CD) Cotton effects appear at 232 nm and 280 nm and a trough appears at 203 nm in the CD spectrum of human prothrombin fragment-2. The Cotton effects in the region from 230 nm to 300 nm are sensitive to pH, ellipticity values at 232 nm increasing from approximately 300 at pH 2.5 to 1300 (degree-cm/decimole) at neutral pH and finally become negative at high pH values. In contrast to fragment-1, at neutral pH the fragment-2 Cotton effect at 232 nm is insensitive to the presence of 8 mM calcium chloride.


1994 ◽  
Vol 11 (3) ◽  
pp. 145-154 ◽  
Author(s):  
M.M. Bhutani ◽  
Ramesh Kumari

The interfacial properties of zirconium oxide, titanium oxide and bismuth oxide have been studied under various experimental conditions. The oxides were characterized by means of their pH titration curves, intrinsic dissociation constants [pKai*(intr.), i = 1, 2) and point of zero charge (pHpzc) in varying concentrations of KNO3 and Na2CrO4 solutions. The pHpzc values of all the oxide samples tended to shift to higher pH values in the presence of Na2CrO4 solution and remain the same in the presence of KNO3 solution. A mechanism for the sorption process is proposed proceeding through an exchange with surface OH– or H2O groups and is interpreted as a case of specific adsorption.


1984 ◽  
Vol 19 (1) ◽  
pp. 87-100
Author(s):  
D. Prasad ◽  
J.G. Henry ◽  
P. Elefsiniotis

Abstract Laboratory studies were conducted to demonstrate the effectiveness of diffused aeration for the removal of ammonia from the effluent of an anaerobic filter treating leachate. The effects of pH, temperature and air flow on the process were studied. The coefficient of desorption of ammonia, KD for the anaerobic filter effluent (TKN 75 mg/L with NH3-N 88%) was determined at pH values of 9, 10 and 11, temperatures of 10, 15, 20, 30 and 35°C, and air flow rates of 50, 120, and 190 cm3/sec/L. Results indicated that nitrogen removal from the effluent of anaerobic filters by ammonia desorption was feasible. Removals exceeding 90% were obtained with 8 hours aeration at pH of 10, a temperature of 20°C, and an air flow rate of 190 cm3/sec/L. Ammonia desorption coefficients, KD, determined at other temperatures and air flow rates can be used to predict ammonia removals under a wide range of operating conditions.


1969 ◽  
Vol 47 (8) ◽  
pp. 1269-1273 ◽  
Author(s):  
A. L. Beauchamp ◽  
J. Israeli ◽  
H. Saulnier

Cu(II), Ni(II), Co(II), and Zn(II) nitrilotriacetates (MeX−) react with histamine nitrate (LH+) to form a protonated mixed complex MeXLH where the metal appears to be bound only to the tertiary imidazolic nitrogen of histaminium ion. At higher pH values the proton dissociates to yield a mixed complex ion MeXL− in which both the imidazolic nitrogen and the terminal amino group are coordinated. The formation constants of these species were calculated from the potentiometric titration curves.


Soil Research ◽  
1981 ◽  
Vol 19 (1) ◽  
pp. 93 ◽  
Author(s):  
GP Gillman

The cation exchange capacity of six surface soils from north Queensland and Hawaii has been measured over a range of pH values (4-6) and ionic strength values (0.003-0.05). The results show that for variable charge soils, modest changes in electrolyte ionic strength are as important in their effect on caton exchange capacity as are changes in pH values.


1991 ◽  
Vol 275 (3) ◽  
pp. 767-773 ◽  
Author(s):  
Y K Li ◽  
J Boggaram ◽  
L D Byers

Two new alkylating reagents, chloro- and bromo-acetylphosphonate, were found to be very effective thiol-blocking reagents. The pH-dependence of the reaction of BAP with 2,4-dinitrothiophenol (25 degrees C, I 0.5) shows a tailing bell-shaped curve (with a plateau at high pH) characteristic of two ionizing groups: the thiol group (pKa 3.2) and the phosphonate group (pKa2 4.6). The rate constant for the reaction of the monoanionic inhibitor with dinitrothiophenolate (k2 = 7 M-1.s-1) is 120 times larger than that of the dianionic species. The haloacetylphosphonates were found to be irreversible inhibitors of glyceraldehyde-3-phosphate dehydrogenase from a variety of sources. They react with the active-site thiol group (Cys-149) and are half-site reagents with yeast glyceraldehyde-3-phosphate dehydrogenase. Thus, when two of the identical four subunits are modified the enzyme is catalytically inactive. The effects of pH (7-10), 2H2O and NAD+ on the reaction with the yeast enzyme were examined in detail. NAD+ enhances the alkylation rates. The second-order rate constant does not show a simple sigmoidal dependence on pH but rather a tailing bell-shaped curve (pKa 7.0 and 8.4) qualitatively similar to that obtained with dinitrothiophenol. There is no significant solvent isotope effect on the limiting rate constants and a normal isotope effect on the two pKa values. The results are consistent with the more reactive enzyme species containing a thiolate and an acidic group that may either donate a proton to the dianionic haloacetylphosphonate or orient the inhibitor.


1974 ◽  
Vol 143 (3) ◽  
pp. 775-777 ◽  
Author(s):  
John L. Wood

The pH-dependence of the degree of hydrogen-bonding between a base and its conjugate acid is considered. When only a small proportion of the total base is complexed, the amount complexed is proportional to (1+coshp)−1 where p=2.303 (pKa–pH), pKa being the dissociation constant of the conjugate acid. This represents sharp pH-dependence. As the proportion complexed increases, the curve broadens, eventually becoming flat-topped, with more than half the base complexed over the range of pH values pKa±logKC, approximately. (K is the complex association constant and C is the formal base concentration, including all forms.) There are similarities to the extent of mono-protonation of a dibasic acid.


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