scholarly journals The effect of Na+ and K+ on the thermal denaturation of Na+ and + K+-dependent ATPase

1983 ◽  
Vol 211 (3) ◽  
pp. 771-774 ◽  
Author(s):  
T H Fischer

To increase our understanding of the physical nature of the Na+ and K+ forms of the Na+ + K+-dependent ATPase, thermal-denaturation studies were conducted in different types of ionic media. Thermal-denaturation measurements were performed by measuring the regeneration of ATPase activity after slow pulse exposure to elevated temperatures. Two types of experiments were performed. First, the dependence of the thermal-denaturation rate on Na+ and K+ concentrations was examined. It was found that both cations stabilized the pump protein. Also, K+ was a more effective stabilizer of the native state than was Na+. Secondly, a set of thermodynamic parameters was obtained by measuring the temperature-dependence of the thermal-denaturation rate under three ionic conditions: 60 mM-K+, 150 mM-Na+ and no Na+ or K+. It was found that ion-mediated stabilization of the pump protein was accompanied by substantial increases in activation enthalpy and entropy, the net effect being a less-pronounced increase in activation free energy.

1969 ◽  
Vol 15 (10) ◽  
pp. 1145-1150 ◽  
Author(s):  
P-C. Tai ◽  
H. Jackson

Several mutants with elevated maximal growth temperature (MGT) were developed from an obligate psychrophile, Micrococcus cryophilus ATCC 15174, by ultraviolet irradiation. Two of the mutants, T8 and M19, have the most similar characteristics to those of their parent. The mutants lost the ability to grow well at 0 °C and showed changes in metabolic pathways while acquiring the ability to grow at elevated temperatures. Heat resistance and deoxyribonucleic acid thermal denaturation temperature were shown to be unrelated to maximum growth temperature. The significance of the mutants is discussed.


2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Hao-Chun Chiang ◽  
Rastislav Levicky

The ability of different crosslinkers to crosslink nanometer thick films of the polymer poly(mercaptopropyl)methylsiloxane (PMPMS), thus stabilizing these films on solid supports, was investigated. The four crosslinkers included 1,11-bismaleimidotriethyleneglycol (BM(PEG)3), tris-(2-maleimidoethyl)amine (TMEA), bismaleimidohexane (BMH), and 1,1′-(methylenedi-4,1-phenylene) bismaleimide (BMDPM). PMPMS films treated with the four crosslinkers were compared in the effectiveness of achieved crosslinking, continuity and stability of the films to rearrangement at elevated temperatures, and modification with single-stranded DNA. The results of electrochemical analyses show that more hydrophilic crosslinkers had difficulty reacting fully with PMPMS thiols, even in these nanometer thin layers. This observation highlights the critical importance of selecting crosslinkers that are chemically compatible. Optimal selection of crosslinker yielded films in which the polymer film was largely incapable of rearranging, even at elevated temperatures, yielding reproducible and stable layers. These results validate use of these supports for applications such as monitoring thermal denaturation of immobilized DNA duplexes.


1976 ◽  
Vol 24 (8) ◽  
pp. 901-907 ◽  
Author(s):  
J D Hunter ◽  
A J Bodner ◽  
F T Hatch ◽  
R L Balhorn ◽  
J A Mazrimas ◽  
...  

The aim of this study was to compare the sensitivity of chromatin from representative cellular stages of spermiogenesis to a single-strandeded nuclease after heat denaturation. Thermal denaturation of chromatin was assayed in situ in fixed round, elongating and elongated spermatids and in testicular sperm from mice. Production of single-stranded deoxyribonucleic acid (DNA) at elevated temperatures was monitored by digesting chromatin with endonuclease specific for single-stranded DNA (S1 nuclease), staining the residual DNA with gallocyanin-chrome alum (GAC) and measuring the stain content by absorption cytophotometry. Changes in GCA staining were minimal over the temperature range of 22-90 degrees C in each cell type not exposed to nuclease. Staining of undigested cells decreased progressively with advancing cell maturity. Nuclease had no effect on the GCA content of round spermatids below 60 degrees C, but above this temperature there was a progressive decrease in GCA-stainable chromatin. Both round and elongating spermatid stages showed a significantly greater sensitivity to nuclease digestion than did more mature stages; sperm showed no effects of nuclease action below 80 degrees C. Progressive chromatin condensation and a concomitant decrease in the number of available DNA phosphate groups during spermiogenic cell maturation may be responsible for the observed decline in sensitivity to nuclease and decreased GCA staining. Thermal denaturation of round spermatids labeled with 3H-thymidine produced no change in autoradiographic mean nuclear grain counts, indicating no loss of thymidine-labeled DNA from the slides during denaturation. When round spermatids and sperm were hydrolyzed with hot tricholoroacetic acid before staining, both nuclear GCA content and autoradiograph grain count were partially reduced, indicating incomplete DNA removal. Almost complete loss of Feulgen-stainable material occurred in these cells and may be due to depurination and elimination of Feulgren-reactant aldehyde groups.


2015 ◽  
Vol 80 (5) ◽  
pp. 613-625 ◽  
Author(s):  
Brankica Raskovic ◽  
Nikolina Babic ◽  
Jelena Korac ◽  
Natalija Polovic

Papain is a protease that consists of ?-helical and ?-sheet domains which unfold almost independently. Both, papain considerable thermal stability and sodium dodecyl sulphate (SDS) resistance have been shown. However, the ability of each domain to unfold upon thermal and SDS denaturation has never been studied. This work shows that fruit papain has slightly higher thermal inactivation resistance when it is compared to stem papain with rather high activation energy (Ea) of 223 ? 16 kJmol-1 and Tm50 value of 79 ? 2 ?C. SDS resistance of fruit papain was estimated by SDS-PAGE analysis and activity staining. It has been noted that, in the presence of SDS, unless heat energy was applied in order to unfold papain, the protein remained active. Furthermore, it has been proven via Fourier transform infrared spectroscopy (FT-IR) that ?-helical domain of fruit papain is more prone to unfolding at elevated temperatures and in the presence of SDS then ?-sheet rich domain. Thermal denaturation of papain without detergent present led to accelerated formation of aggregation specific intermolecular ?-sheets as compared to native protein. Presented results are both, of fundamental and application importance.


1979 ◽  
Vol 44 (10) ◽  
pp. 3090-3101
Author(s):  
Eduard Brynda ◽  
Miroslav Bleha

The reversible thermal denaturation of chymotrypsinogen A bound to the insoluble poly(2-hydroxyethyl methacrylate) Spheron matrix was investigated by the fluorescence method. The applicability of fluorescence data in the calculation of the thermodynamic parameters of denaturation was tested. Equilibrium data were obtained for immobilized chymotrypsinogen and chymotrypsinogen in solution at various pH in the range 2-6. The binding to Spheron shifts the thermodynamic equlibrium in favour of the denaturated state. An increase in pH above 3 did not affect the denaturation of immobilized chymotrypsinogen. The denaturation is controlled by the first-order kinetics. The rate constants and magnitudes of changes of free energy, enthalpy and entropy were calculated for the transition from the native state into the activated state, and from denaturated state into the activated state. The temperature dependence of the rate constants for the denaturation of immobilized chymotrypsinogen is qualitatively different from for chymotrypsinogen in solution. The results were interpreted as effects of the physical interaction between the denaturated protein and polymeric matrix.


2012 ◽  
Vol 9 (9) ◽  
pp. 3531-3545 ◽  
Author(s):  
J. Taucher ◽  
K. G. Schulz ◽  
T. Dittmar ◽  
U. Sommer ◽  
A. Oschlies ◽  
...  

Abstract. Increasing concentrations of atmospheric carbon dioxide are projected to lead to an increase in sea surface temperatures, potentially impacting marine ecosystems and biogeochemical cycling. Here we conducted an indoor mesocosm experiment with a natural plankton community taken from the Baltic Sea in summer. We induced a plankton bloom via nutrient addition and followed the dynamics of the different carbon and nitrogen pools for a period of one month at temperatures ranging from 9.5 °C to 17.5 °C, representing a range of ±4 °C relative to ambient temperature. The uptake of dissolved inorganic carbon (DIC) and the net build-up of both particulate (POC) and dissolved organic carbon (DOC) were all enhanced at higher temperatures and almost doubled over a temperature gradient of 8 °C. Furthermore, elemental ratios of carbon and nitrogen (C : N) in both particulate and dissolved organic matter increased in response to higher temperatures, both reaching very high C : N ratios of > 30 at +4 °C. Altogether, these observations suggest a pronounced increase in excess carbon fixation in response to elevated temperatures. Most of these findings are contrary to results from similar experiments conducted with plankton populations sampled in spring, revealing large uncertainties in our knowledge of temperature sensitivities of key processes in marine carbon cycling. Since a major difference to previous mesocosm experiments was the dominant phytoplankton species, we hypothesize that species composition might play an important role in the response of biogeochemical cycling to increasing temperatures.


1954 ◽  
Vol 27 (3) ◽  
pp. 648-670
Author(s):  
Arturo Chiesa

Abstract The T-50 test, which has already been used for the last twenty years in many rubber factories has, up to the present time, been used solely as a means of checking vulcanization conditions, and so far no one has attempted to derive any mathematical relations from the results obtained. In the first part of this work it is shown that the T-50 test can be regarded as a useful and efficient means for studying much more complex and important problems, as, for example, the determination of the vulcanization characteristics of a rubber compound, both with respect to the ingredients and from the thermal point of view. In fact, this test makes it possible to obtain quantative data rapidly, which can be utilized to render any study easier and more conclusive than is possible with other tests commonly used, e.g., dynamometric parameters, aging tests, relaxation at elevated temperatures, etc. In making a study of the T-50 test, the approach was from the point of view of the chemical and physical nature of the phenomena involved in the test. It has been established with a considerable degree of exactitude that results obtained with the test conform to an energy law common to many chemical-physical phenomena, viz., the law of Arrhenius, which expresses the relation between the rate of a reaction and the temperature of a process by means of a parameter which depends on the activation energy of the process itself. In fact, analysis of experimental data shows clearly that, except in the case of mixtures having peculiar vulcanization characteristics, the activation energy of the process is practically independent of the composition of the mixture. One is led to believe, therefore, that such energy depends directly on the nature of the polymer itself and on its vulcanization reactions with sulfur, and that it is independent of the chemical and physical factors which control vulcanization reactions. On account of the nature of our work, we could not undertake a thorough scientific study of the aspect of this fact, considered from the point of view of the molecular structure of cross-linked high polymers. Instead, we have limited ourselves to developing an application of considerable practical utility, based on the fact that the activation energy is practically constant. In fact, a simple correlation diagram can be established which can be used for most types of vulcanizates, and by means of which it is possible to obtain directly the equivalent coefficients for passing from one vulcanization temperature to another with the longest temperature range which it was possible to use, i.e., from 78° to 151 ° C. It has thus been possible to interpret the results of the T-50 test for temperatures other than for the temperature at which the test was made, and thus to obtain a more complete and comprehensive picture directly related to the same chemico-physical phenomena. Passing on to the subject of dynamometric parameters, it is shown that, for these too, the above law applies with sufficient approximation. Again, since the order of magnitude of the activation energy is similar to that found by the T-50 test, the same correlation diagram applies, thus appreciably increasing its practical importance. Finally, the satisfactory accord between the activation energy values obtained by the T-50 test and those obtained by stress relaxation tests reveals an intimate relation between the two phenomena, which otherwise would appear to be completely different. Thus a further contribution is made to the complex study of relaxation.


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