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1.
Differential scanning calorimetry and electron paramagnetic resonance experiments were performed on glycerinated skeletal muscle fibres to study the effect of the binding of nucleotides and nucleotide analogues to myosin. The thermal unfolding of muscle fibres in rigor showed three discrete domain regions with thermal stability of 52.2, 58.8 and 67.8°C. AMP.PNP and ATP plus AlF3 or BeF2 affected markedly the transitions, which implies the strong interaction between AMP.PNP or nucleotide analogues and catalytic domain of myosin, and a partial dissociation of heads from actin. ADP.BeFx and states model the transition states of the ATP hydrolysis cycle which precede the powerstroke of the muscle fibres. Spectrum deconvolution on isothiocyanate-labelled fibres in AMP.PNP-state resulted in two populations; 50% of labels was highly ordered with respect to fibre axis, whereas the other 50% of labels was randomly oriented. The myosin heads which showed high degree of order were in the strongly binding ADP-state. The spectra in - and ADP.BeFx state reflected random orientation of labels with increased rotational mobility in comparison with rigor. The results suggest that myosin in muscle fibres in ADP.BeFx state exists in two forms. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

2.
Thermal stability and internal dynamics of myosin head in psoas muscle fibres of rabbit in the intermediate state AM.ADP.Pi - mimicked by AM.ADP.Vi - of the ATP hydrolysis cycle was studied by differential scanning calorimetry and spin label electron paramagnetic resonance spectroscopy. Three overlapping endotherms were detected in rigor, in strongly binding ADP and weakly binding AM.ADP.Vi state of myosin to actin. The transition at 54.0°C can be assigned to the 50 k actin-binding domain. The transition at highest temperature (67.3°C) represents the unfolding of actin and the contributions arising from the nucleotide-myosin head interaction. The transition at 58.4°C reflects the melting of the large rod part of myosin. Nucleotide binding (ADP, ATP plus orthovanadate) induced shifts of the melting temperatures and produced changes in the calorimetric enthalpies. The changes of the EPR parameters indicated local rearrangements of the internal structure in myosin heads in agreement with DSC findings. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

3.
The internal dynamics and the thermal stability of myosin in rabbit psoas muscle fibres in different intermediate states of the ATP hydrolysis cycle were studied by differential scanning calorimetry (DSC) and electron paramagnetic resonance (EPR) spectroscopy. Three overlapping endotherms were detected in rigor, in strongly binding and weakly binding state of myosin to actin. The transition at 58.4°C can be assigned to the nucleotide-binding domain. The transition at highest temperature represents the unfolding of the actin and the contributions arising from the actin-myosin interaction. The transition of 54°C reflects the interaction between the subunits of myosin. Nucleotide binding induced shifts of the melting temperatures and produced variations in the calorimetric enthalpy changes. The changes of the EPR parameters indicated local rearrangements of the internal structure in myosin heads. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

4.
Differential scanning calorimetry and electron paramagnetic resonance experiments were performed on glycerinated muscle fibres to study the effect of the binding of nucleotides (ADP and AMP⋅PNP) to myosin. The thermal unfolding of muscle fibres showed three discrete domain regions with thermal stabilities of 52.2, 58.8 and 67.8°C. AMP⋅PNP markedly affected the transitions, implying the strong interaction between AMP⋅PNP and catalytic domain, and partial dissociation of heads from actin. ADP produced only small changes in transition temperatures. Spectrum deconvolution performed on isothiocyanate-labelled fibres in AMP⋅PNP-state resulted in two populations; 50% of labels was highly ordered with respect to fibre axis, whereas the other 50% of labels was randomly oriented. The conformation of the myosin heads which showed high degree of order were in the strongly binding ADP-state, the heads being attached to actin differ from those of heads in rigor. The results support the suggestion that the attached heads in strongly binding state to actin might have different local conformations. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

5.
Electron paramagnetic resonance (EPR, ST-EPR) and differential scanning calorimetry(DSC) were used in conventional and temperature modulated mode to study internal motions and energetics of myosin in skeletal muscle fibres in different states of the actomyosin ATPase cycle. Psoas muscle fibres from rabbit were spin-labelled with an isothiocyanate-based probe molecule at the reactive sulfhydryl site (Cys-707) of the catalytic domain of myosin. In the presence of nucleotides (ATP, ADP, AMP⋅PNP) and ATP or ADP plus orthovanadate, the conventional EPR spectra showed changes in the ordering of the probe molecules in fibres. In MgADP state a new distribution appeared; ATP plus orthovanadate increased the orientational disorder of myosin heads, a random population of spin labels was superimposed on the ADP-like spectrum. In the complex DSC pattern, higher transition referred to the head region of myosin. The enthalpy of the thermal unfolding depended on the nucleotides, the conversion from a strongly attached state of myosin to actin to a weakly binding state was accompanied with an increase of the transition temperature which was due to the change of the affinity of nucleotide binding to myosin. This was more pronounced in TMDSC mode, indicating that the strong-binding state and rigor state differ energetically from each other. The different transition temperatures indicated alterations in the internal microstructure of myosin head region The monoton decreasing TMDSC heat capacities show that C p of biological samples should not be temperature independent. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

6.
Actin is one of the main components in the eukaryote cells which plays significant role in many cellular processes, like force-generation, maintenance of the shape of cells, cell-division cycle and transport processes. In this study the thermal transitions of monomer and polymerized actins were studied to get information about the changes induced by polymerization and binding of myosin to actin using DSC and EPR techniques. The main thermal transition of F-actin was at 67.5°C by EPR using spin-labeled actin (the relative viscosity change was around 62°C), while the DSC denaturation T ms were at 60.3d°C for G-actin and at 70.5°C for F-actin. Applying the Lumry-Eyring model to obtain the parameters of the kinetic process and calculate the activation energy, a ‘break’ was found for F-actin in the function of first-order kinetic constant vs. 1/T. This indicates that an altered interdomain interaction is present in F-actin. The addition of myosin or heavy meromyosin (HMM) in different molar ratio of myosin to actin has changed significantly the EPR spectrum of spin-labeled F-actin, indicating the presence of the supramolecular complex. Analyzing the DSC traces of the actomyosin complex it was possible to identify the different structural domains of myosin and actin.  相似文献   

7.
Summary The heat capacity of contractile proteins actin and myosin was studied in psoas muscle of rabbit in strongly and weakly binding state of myosin to actin as a function of temperature by DSC. Deconvolution of the unfolding scans makes possible to characterize the structural domains of the macromolecules. We tried to approach the unfolding process in different intermediate state of ATP hydrolysis. The thermal transitions were calorimetrically irreversible, therefore the two-state irreversible model that describes fairly well the denaturation of different proteins was used for evaluation of the denaturation processes in muscle fibers in strongly (rigor, ADP) and weakly binding states (ATP·Vi, ADP·AlF4) of myosin to actin. Deconvolution resulted in four transitions, the first three transition temperatures were almost independent of the intermediate states of muscle, the last transition temperature was shifted to higher temperature, when the buffer solution was manipulated. The mean values in strongly binding states were Tm1=52.9±0.7°C, Tm2=57.9±0.7°C, Tm3=63.7±1.0°C and Tm4=67.8±0.7°C, but the last transition increased to higher temperature depending on the Pi analogue.  相似文献   

8.
Differential scanning calorimetry (DSC) and electron paramagnetic resonance spectroscopy (EPR, both conventional and saturation transfer EPR) were used to study the motional dynamics and segmental flexibility of myosin in muscle fibres in the presence of free radical generating system. Muscle fibre bundles isolated from psoas muscle of rabbit were spin-labelled with maleimide- and isothiocyanate-based probe molecules at the reactive sulfhydryl sites (Cys-707) of the motor domain. In the presence of hydroxyl free radicals the spectral intensity of the maleimide probe molecules decreased with time following a single exponential curve. MgADP and MgATP plus orthovanadate that produce flexibility changes in the multisubunit structure of myosin enhanced the reduction of the attached nitroxide molecules in free radical generating system. The analysis of the EPR spectra of spin-labelled and oriented fibres showed that the narrow distribution of spin labels changed in the presence of hydroxyl free radicals. Spectrum analysis by computer subtraction showed that short irradiation by UV light resulted in the enhancement of the ordered population at the expense of the disordered population. This suggests a transition of myosin heads from weak- binding state into strong-binding state. DSC measurements performed on calf cardiac myosin resulted in two main transitions at 49.4 and 54.1°C, respectively. Addition of MgADP produced a decrease of the 49.4°C transition, whereas a shift towards higher temperature was detected at the 54.1°C transition. It shows that there is an inter-site communication between the domains of the myosin. Hydroxyl free radicals induced further shifts of the transition temperatures and affected the width of the heat absorption curves. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

9.
PMR-15 is a high-performance thermoset polyimide resin that is used in many high-temperature applications. Postcured PMR-15 produces room-temperature electron paramagnetic resonance (EPR) spectra from stable freeradical species that are formed during the postcuring stages. The variable-temperature EPR spectral intensities show a minimum at Tmin in the range ?60 to ?40°C, and a maximum at Tmax in the range 80–120°C. The EPR intensities follow the inverse temperature dependence of Curie's law below Tmin and are due to a stable free radical. The intensities then increase with increasing temperature between Tmin and Tmax. The free radical with such temperature dependence is not present below Tmin and is undetectable by EPR at temperatures above Tmax. These free radicals are generated during the postcuring process at elevated temperature above 310°C. The thermo-oxidative degradation involves free radicals generated during the postcuring process in the presence of oxygen gas. © 1993 John Wiley & Sons, Inc.  相似文献   

10.
Transition temperature and thermal stability of proteins were studied in the presence and absence of boron. The observed midpoint of thermal denaturation (T m) of cytochrome c (Cyt c) at pH 9.2 was 68.8 °C, which in the presence of boron increased to 71.0 °C. For metmyoglobin, T m increased from 79.7 °C in the absence of boron to 83.5 °C in the presence of boron. Boron caused an increase of 10% in the reversibility of thermal denaturation of cytochrome c when compared with control. Activity measurements of the heat treated proteins and T m suggest an increased thermal stability toward inactivation and denaturation of heme proteins in the presence of boron.  相似文献   

11.
A phase diagram of the tetraethylammonium hydroxide-water binary system was studied by differential thermal analysis. Seven crystal phases of tetraethylammonium hydroxide were found. Two of them are stable: tetrahydrate (T m = 47.0°C), two phases of a compound with the hydration number 7.5 (T m = −18.1 and −23.2°C); five are metastable phases: two polymorphs of pentahydrate (T m = 39.4 and 0.9°C), hexahydrate (T m = −70.1°C), and a compound with hydrate number 10–11 (T m = −112°C).  相似文献   

12.
Poly(dicyanoacetylene) (PDCA) has been synthesized and characterized. The pristine polymer has EPR g-value, linewidth, unpaired spin concentration, spin—spin relaxation time (T2), and room temperature dc conductivity (σRT) very similar to those of pristine cis-polyacetylene (PA), but shorter spin—lattice relaxation time (T1). Saturation doping with iodine has little effect on most EPR characteristics of the polymer except for a slight increase in T1. The doped PDCA has σRT value of only 5 X 10-9 (Ω cm)-1, indicating either low carrier concentration and/or carrier mobility. Partial cyclization of the nitrile groups by heating at 400°C of PDCA produces l-PDCA with significant increases in unpaired spin concentration and σRT but marginal effects on other properties. Saturation doping of l-PDCA with iodine increases σRT to 7 × 10-3 (Ω cm)-1 without appreciable changes in EPR characteristics. The dopants in both polymers can be removed by evacuation indicating only weak charge transfer interactions. The possible stereoelectronic contribution toward the property differences between the PDCA polymers and PA are discussed.  相似文献   

13.
The effect of butyl acrylate (BA), divinyl benzene (DVB) and vinyltrimethoxysilane (TMVS) on the thermal properties of poly(methyl methacrylate-co-butyl acrylate-co-acrylic acid) was investigated. Glass transition temperature (Tg), melting temperature (Tm) and specific heat capacity of the copolymers were investigated using Differential Scanning Calorimetry. Thermal stability of the copolymers which is associated with the degradation temperature (Td) was studied by Thermogravimetric Analysis. Polyacrylates with Tg ranges between -19°Cand 19°C were obtained. With the incorporation of >7 wt% of DVB, the Tg of the copolymer increases from about ?17°C to ?10°C even though they have not undergone UV irradiation. Gel content results prove that crosslinking has occurred in the copolymers. With increasing amount of TMVS from 0 wt% to 7 wt%, the Tm of the copolymers prepared at acidic pH is about 40-60°C higher than that at the alkaline pH. However, the addition of TMVS gives no significant effect to the Tg and Td of the copolymer films. The thermal stability of the copolymer has improved with increasing amount of BA and DVB, with DVB being more effective. The highest Td of 425°C with 8% of DVB has been obtained. Consequently, a polyacrylate copolymer with a Tg of about ?13°C, a Tm of 170 °C and a Td of about 424°C has been successfully synthesized. Hence, the soft polyacrylate with its relatively high Tm and Td could serve as a superb material especially to be applied in the areas that require high melting temperature and good thermal stability.  相似文献   

14.
Poly[2,2-bis(4-oxycyclohexyl)propane adipate] was formed by melt polymerizing adipic acid and 2,2-bis(4-hydroxycyclohexyl)propane in the presence of n-butylhydroxyoxostannane. This polyester possessed a melting temperature (Tm) of 200°C and a glass transition temperature (Tg) of 75°C. An excellent notched impact strength of 22 ft. lbs./in. was its best physical property.  相似文献   

15.
A series of fully aromatic, thermotropic homopolyesters, derived from 3,4′-benzophenone dicarboxylic acid and various aromatic diols, was prepared by the melt polycondensation method and examined for thermotropic behavior by a variety of experimental techniques. The aromatic diols used in the study were hydroquinone, 2,6-, 1,4-, 1,5-, 2,3-, and 2,7-naphthalenediol isomers. All of the homopolyesters of 3,4′-benzophenone dicarboxylic acid with aromatic diols (except that with 2,7-naphthalenediol) formed a nematic LC phase in the melt. They had the glass transition temperatures (Tg) in the range of 133–164°C, the melting transitions (Tm) in the range 305–360°C and the high thermal stabilities (Td) in the range of 410–483°C. The 2,6-naphthalenediol based homopolymer had the highest Tm (360°C) and the 2,3-naphthalenediol based homopolymer had the lowest Tm (305°C) among all of the homopolymers of naphthalenediol isomers. © 1994 John Wiley & Sons, Inc.  相似文献   

16.
Annealing of poly(butylene terephthalate) (PBT) was studied by differential scanning calorimetry (DSC) and small angle X‐ray scattering (SAXS) measurement. A PBT sample was annealed at a recrystallization temperature where recrystallization occurs with a maximum rate in the heating process of the sample. In the subsequent annealing steps, the annealed sample was annealed repeatedly at the recrystallization temperatures, and the stepwise annealing sample was obtained. Peak melting temperature (Tm) and sharpness of DSC peak of the stepwise annealing sample increased with the annealing step. A high melting‐temperature sample was obtained in a short time, and Tm increased up to 238.5°C which is higher than all the Tm values that appear in the literature. The long period calculated from SAXS curves of the stepwise annealing sample increased with the annealing step. The increase of crystallite size and perfection of the crystal in the stepwise annealing process is suggested. Annealing experiment indicated that T°m should be higher than about 235°C. Tm increased linearly with the annealing temperature of the final step in the stepwise annealing (Ta). The equilibrium melting temperature (T°m) for PBT was estimated to be 247°C by the application of a Hoffman–Weeks plot to the relation between Tm vs. Ta. © 1999 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 37: 2420–2429, 1999  相似文献   

17.
The esterification of poly(γ-glutamic acid) (γ-PGA) produced by Bacillus subtilis F2-01 with alkyl halides was carried out at 60°C in N-methyl-2-pyrrolidinone (NMP) in the presence of sodium bicarbonate to obtain the corresponding esterified γ-PGA. The thermal properties of these γ-PGA esters were examined by differential scanning calorimetry and thermogravimetry. γ-PGA esters were more stable than free acid type γ-PGA, which decomposed at 210°C. Melting temperature (Tm) of γ-PGA esters could be observed at 230-250°C. Tm of γ-PGA n-alkyl esters reached a maximum at an alkyl chain length of n = 3. © 1995 John Wiley & Sons, Inc.  相似文献   

18.
A new α-amylase was extracted from a recently found strain of Bacillus sp. and purified by ion-exchange chromatography. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed a single band for the purified enzyme with an apparent molecular weight of 59 kDa. The optimum temperature and pH range of the enzyme were 40–60°C and 4.5–7.5, respectively, and its activation energy was 1.974 kcal/mol. The K m value for the enzyme activity on solubie starch was 4 mg/mL, and the T m values obtained from the circular dichroism (CD) results of thermal unfolding were 78.7 and 80.2°C in the absence and presence of the calcium, respectively. The enzyme was almost completely inhibited by the addition of Fe3+, Mn2+, and Zn2+ and was activated by EDTA, Cr3+, and Al3+. Moreover, it was partially inhibited by Ca2+, Ba2+, Ni2+, and Co2+. Proteolytic digestion of the enzyme using trypsin combined with results from T m using CD and irreversible thermoinactivation suggests that this enzyme can be considered a moderate thermophile with both mild flexibility and rigidity.  相似文献   

19.
The thermal stability of a short carbon-fiber-reinforced PEEK composite was assessed by thermogravimetry and by a Rheometrics dynamic analyzer. The results indicated that holding for 10 min at 380°C was a suitable melting condition to avoid the thermooxidative degradation under air. After proving that the heating rate of 50°C/min can be used to evaluate the crystallinity, a heating stage was used to prepare nonisothermally crystallized specimens using cooling rates from 1 to 100°C/min after melting at 400°C for 3 or 15 min. The degree of crystallinity and the melting behavior of these specimens were investigated by DSC at a heating rate of 50°C/min. The presence of three or four regions indicated that the upper melting temperature, Tm, changed with the crystallization temperature. The first region with the highest Tm, which corresponded to the cooling rate of 1°C/min, can be associated with the crystallization in regime II. There was a second region where Tm decreased as the amount of crystals formed in regime II decreased with increasing cooling rate from 5 to 20°C/min. The third region, a plateau region, corresponded to regime III condition in which the crystals were imperfect. In the fourth region, the cooling was so fast that crystallization was incomplete during the cooling for the melting condition of 400°C for 15 min. © 1998 John Wiley & Sons, Inc. J. Polym. Sci. B Polym. Phys. 36: 2225–2235, 1998  相似文献   

20.
A novel phosphorus-containing bismaleimide, 3,3′-bis(maleimidophenyl)phenylphosphine oxide (BMPPPO), was synthesized from triphenylphosphine oxide. This bismaleimide exhibited good solubility in common organic solvents, such as methylethylketone, methylisobutylketone, dichloromethane, chloroform, tetrahydrofuran, acetone, methanol, ethanol, and hot toluene. A low melting point (Tm = 148 °C), a relatively low polymerization temperature (Tp = 214 °C), and a wide processing window (TpTm = 66 °C) were also obtained for BMPPPO. This implies better processing capability. In contrast to most known phosphorus-containing polymers, the incorporation of BMPPPO into poly(bismaleimide) enhanced the polymer glass-transition temperature. Thermal stability at temperatures over 550 °C and char yields in the high-temperature region over 700 °C were also improved. As a result, the flame-retardant properties of the poly(bismaleimide)s were improved. © 2001 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 39: 1716–1725, 2001  相似文献   

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