Temperature-dependent X-ray diffraction as a probe of protein structural dynamics

Frauenfelder, Hans; Petsko, Gregory A.; Tsernoglou, Demetrius

doi:10.1038/280558a0

Cited by 1,163 publications

(664 citation statements)

References 28 publications

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“…The samples were contained in aluminium flat-plate cells of 0.5 mm thickness. Neutron scattering intensities were recorded on the following samples: 63 mg xylanase in 100% D 2 Data were collected over the temperature range from 100 K to 300 K for each methanol concentration. The samples were first cooled to B100 K and then slowly reheated in steps back to 300 K over a period of B6 h, during which time the data were recorded.…”

Section: Data Collectionmentioning

confidence: 99%

“…Experimental techniques such as Mo¨ssbauer spectroscopy, X-ray diffraction and neutron scattering have suggested the presence of a transition in the internal dynamics of protein motions at B180-220 K. [1][2][3][4][5][6][7][8][9] This apparent dynamical transition has been observed to have similarities to the phase transition that appears in glass-forming liquids. [10][11][12][13] The motions below the transition are believed to be mostly harmonic whereas the mean-square displacements above the transition are dominated by anharmonic contributions.…”

Section: Introductionmentioning

confidence: 99%

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Temperature and timescale dependence of protein dynamics in methanol : water mixtures

Tournier

Réat

Dunn

et al. 2005

Phys. Chem. Chem. Phys.

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Experimental and computer simulation studies have suggested the presence of a transition in the dynamics of hydrated proteins at around 180-220 K. This transition is manifested by nonlinear behaviour in the temperature dependence of the average atomic mean-square displacement which increases at high temperature. Here, we present results of a dynamic neutron scattering analysis of the transition for a simple enzyme: xylanase in water : methanol solutions of varying methanol concentrations. In order to investigate motions on different timescales, two different instruments were used: one sensitive to B100 ps timescale motions and the other to Bns timescale motions. The results reveal distinctly different behaviour on the two timescales examined. On the shorter timescale the dynamics are dictated by the properties of the surrounding solvent: the temperature of the dynamical transition lowers with increasing methanol concentration closely following the melting behaviour of the corresponding water : methanol solution. This contrasts with the longer (ns) timescale results in which the dynamical transition appears at temperatures lower than the corresponding melting point of the cryosolvent. These results are suggested to arise from a collaborative effect between the protein surface and the solvent which lowers the effective melting temperature of the protein hydration layer. Taken together, the results suggest that the protein solvation shell may play a major role in the temperature dependence of protein solution dynamics.

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Section: Data Collectionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Temperature and timescale dependence of protein dynamics in methanol : water mixtures

Tournier

Réat

Dunn

et al. 2005

Phys. Chem. Chem. Phys.

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“…There is considerable evidence that enzyme activity is dependent upon enzyme flexibility [53][54][55][56]. It is implicit in the concept of induced fit [57,58] that an enzyme must flex over a time scale in keeping with its catalytic-centre activity, and that changes between conformational substates allow catalytic activity.…”

Section: Inter-relationship Of Enzyme Stability and Activitymentioning

confidence: 99%

“…However, excessive flexibility is the first step towards denaturation, so although an enzyme must be sufficiently rigid to have a reserve of stability, it must also have the flexibility needed for effective catalysis. These are of course generalizations, since stability is a global property of an enzyme [25] whereas the flexibility required for activity may well be localized [54][55][56]. They do, however, explain why enzymes tend in general to be denatured at temperatures not very far above their ' design ' temperature.…”

Section: Inter-relationship Of Enzyme Stability and Activitymentioning

confidence: 99%

The denaturation and degradation of stable enzymes at high temperatures

Daniel

Dines

Petach³

1996

Biochemical Journal

271

175

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Now that enzymes are available that are stable above 100 mC it is possible to investigate conformational stability at this temperature, and also the effect of high-temperature degradative reactions in functioning enzymes and the inter-relationship between degradation and denaturation. The conformational stability of proteins depends upon stabilizing forces arising from a large number of weak interactions, which are opposed by an almost equally large destabilizing force due mostly to conformational entropy. The difference between these, the net free energy of stabilization, is relatively small, equivalent to a few interactions. The enhanced stability of very stable proteins can be achieved by an additional stabilizing force which is again equivalent to only a few stabilizing interactions. There is currently no strong evidence that any particular interaction (e.g. hydrogen bonds, hydrophobic interactions) plays a more important role in

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“…For example, 0006-2960/88/0427-2132$01.50/0 a side chain may have rotated, some hydrogen bonds may have shifted, a single helix may be displaced, or small movements may occur involving entire structural domains (Austin et al, 1975;Frauenfelder et al, 1979;Hartmann et al, 1982;Ringe et al, 1985). Recently, a hierarchical model for such substates has been proposed, each level of the hierarchy being characterized by a specific interconversion time scale and energy (Ansari et al, 1985(Ansari et al, , 1987Frauenfelder & Gratton, 1985;Kuriyan et al, 1987).…”

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confidence: 99%

Effect of unfolding on the tryptophanyl fluorescence lifetime distribution in apomyoglobin

Bismuto¹,

Gratton²,

Irace³

1988

Biochemistry

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Proteins exhibit, even in their native state, a large number of conformations differing in small details (substates). The fluorescence lifetime of tryptophanyl residues can reflect the microenvironmental characteristics of these subconformations. We have analyzed the lifetime distribution of the unique indole residue of tuna apomyoglobin (Trp A-12) during the unfolding induced by temperature or guanidine hydrochloride. The results show that the increase of the temperature from 10 to 30 OC causes a sharpening

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Temperature-dependent X-ray diffraction as a probe of protein structural dynamics

Cited by 1,163 publications

References 28 publications

Temperature and timescale dependence of protein dynamics in methanol : water mixtures

Temperature and timescale dependence of protein dynamics in methanol : water mixtures

The denaturation and degradation of stable enzymes at high temperatures

Effect of unfolding on the tryptophanyl fluorescence lifetime distribution in apomyoglobin

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