Structural stability of actin filaments as studied by DSC and EPR

Lőrinczy, D.; Könczöl, Franciska; Gaszner, Balázs; Belágyi, József

doi:10.1016/s0040-6031(98)00495-x

Cited by 44 publications

(30 citation statements)

References 18 publications

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“…The calorimetric enthalpy attributed to this transition increased in the presence of phalloidin, while jasplakinolide decreased the enthalpy substantially. Considering that in previous studies solution of actin monomers gave T m value of 53.4°C [17][18][19], one may speculate that the actin monomer content of the samples was responsible for the observations. However, in the case of actin concentrations applied here (46 lM) only less than 0.5% of the actin should have been in the monomeric form considering the critical concentration for monomer assembly ($0.1 lM), and thus this monomer content appears to be much less than enough to account for the low temperature transitions.…”

Section: Differential Scanning Calorimetrymentioning

confidence: 63%

The effect of phalloidin and jasplakinolide on the flexibility and thermal stability of actin filaments

VISEGRADY¹

2004

FEBS Letters

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In this work the effect of phalloidin and jasplakinolide on the dynamic properties and thermal stability of actin filaments was studied. Temperature dependent fluorescence resonance energy transfer measurements showed that filaments of Ca-actin became more rigid in the presence of phalloidin or jasplakinolide. Differential scanning calorimetric data implied that the stiffer filaments also had greater thermal stability in the presence of phalloidin or jasplakinolide. The fluorescence and calorimetric measurements provided evidences that the extent of stabilization by jasplakinolide was greater than that by phalloidin.

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Section: Differential Scanning Calorimetrymentioning

confidence: 63%

The effect of phalloidin and jasplakinolide on the flexibility and thermal stability of actin filaments

VISEGRADY¹

2004

FEBS Letters

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“…Comparing the different T m values it is possible to characterize the thermodynamic stability of proteins under different conditions [24,30]. A higher T m value is typically correlated with a thermodynamically more stable protein structure [30,31].…”

Section: Resultsmentioning

confidence: 99%

“…Previous experiments showed that DSC is an effective method to study the thermal properties of actin [15][16][17][18][19][20][21][22][23][24]. The aim of our work was to investigate the effect of jasplakinolide on the thermodynamic properties of the ADP.BeF x bound actin filaments with differential scanning calorimetry (DSC).…”

Section: Introductionmentioning

confidence: 99%

The effect of jasplakinolide on the thermodynamic properties of ADP.BeFx bound actin filaments

et al. 2007

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The effect of BeF x and a natural toxin (jasplakinolide) was examined on the thermal stability of actin filaments by using differential scanning calorimetry. The phosphate analogue beryllium fluoride shifted the melting temperature of actin filaments (67.4 °C) to 83.7 °C indicating that the filaments were thermodynamically more stable in their complex with ADP.BeF x . A similar tendency was observed when the jasplakinolide was used in the absence of BeF x . When both the ADP.BeF x and the jasplakinolide bound to the actin filaments their collective effect was similar to that observed with ADP.BeF x or jasplakinolide alone. These results suggested that ADP.BeF x and jasplakinolide probably stabilize the actin filaments by similar molecular mechanisms.

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“…These values are similar to those previously published by others. 22,24,25,33 For concentrations inferior to 0.15 mg/mL, the exothermic peak produced by aggregation has a very large cooperativity and merges with the baseline. Moreover, a pretransition is present at around 508C for these very low concentrations.…”

Section: Effect Of the Concentration On The Thermograms Of F-actinmentioning

confidence: 99%

“…[18][19][20][21][22][23][24] This was the case for F-actin in which a single peak was deconvoluted into three components, two interpreted as the unfold-ing of independent domains and the third attributed to the interaction between the domains. 25 Actin-ligand interactions and the existence of domains have been studied with DSC, but generally conclusions were drawn using only this technique and from only one scan under a single experimental condition. 24,[26][27][28][29][30][31][32][33] In the present work, we have shown that the endothermic peak of the heat denaturation of F-actin is produced by three events that occur at closely spaced temperatures.…”

Section: Introductionmentioning

confidence: 99%

Three steps in the thermal unfolding of F‐actin: An experimental evidence

Gicquaud

Heppell

2006

Biopolymers

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The development of differential scanning calorimetry has resulted in an increased interest in studies of the unfolding process in proteins with the aim of identifying domains and interactions with ligands or other proteins. Several of these studies were done with actin and showed that the thermal unfolding of F-actin occurs in at least three steps; this was interpreted as the denaturation of independent domains. In the present work, we have followed the thermal unfolding of F-actin using differential scanning calorimetry (DSC), CD spectroscopy, and probe fluorescence. We found that the three steps revealed through DSC are not the denaturation of independent domains. These three steps are a change in the environment of cys 374 at 49.5 degrees C; a modification at the nucleotide-binding site at 55 degrees C; and the unfolding of the peptide chain at 64 degrees C. Previous interpretations of the thermograms of F-actin were thus erroneous. Since DSC is now widely used to study proteins, our experimental approach and conclusions may also be relevant in denaturation studies of proteins in general.

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Structural stability of actin filaments as studied by DSC and EPR

Cited by 44 publications

References 18 publications

The effect of phalloidin and jasplakinolide on the flexibility and thermal stability of actin filaments

The effect of phalloidin and jasplakinolide on the flexibility and thermal stability of actin filaments

The effect of jasplakinolide on the thermodynamic properties of ADP.BeFx bound actin filaments

Three steps in the thermal unfolding of F‐actin: An experimental evidence

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