Glyceraldehyde-3-phosphate dehydrogenase from Thermotoga maritima: Strategies of protein stabilization

Jaenicke, Rainer

doi:10.1016/0168-6445(96)00013-7

Cited by 9 publications

(11 citation statements)

References 23 publications

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“…It has been suggested earlier that thermostable proteins show ‘anomalous rigidity’ . This assumption appears to be in strong contrast to the study by Seewald et al .…”

Section: Heat Capacity Configurational Entropy and Fluctuations In Pcontrasting

confidence: 70%

“…At least in certain cases, thermostable proteins may have unusually high flexibility at high temperatures. The proposal that thermostable proteins are unusually rigid is primarily based on the low H–D exchange rates and low protease susceptibility . However, slower H–D exchange rates, as well as the impediment of protease attack, may be a consequence of the shorter time interval that thermostable proteins spend in non‐native states, which is directly related to their thermal stability.…”

Section: Heat Capacity Configurational Entropy and Fluctuations In Pmentioning

confidence: 99%

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Rigidity versus flexibility: the dilemma of understanding protein thermal stability

2015

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The role of fluctuations in protein thermostability has recently received considerable attention. In the current literature a dualistic picture can be found: thermostability seems to be associated with enhanced rigidity of the protein scaffold in parallel with the reduction of flexible parts of the structure. In contradiction to such arguments it has been shown by experimental studies and computer simulation that thermal tolerance of a protein is not necessarily correlated with the suppression of internal fluctuations and mobility. Both concepts, rigidity and flexibility, are derived from mechanical engineering and represent temporally insensitive features describing static properties, neglecting that relative motion at certain time scales is possible in structurally stable regions of a protein. This suggests that a strict separation of rigid and flexible parts of a protein molecule does not describe the reality correctly. In this work the concepts of mobility/flexibility versus rigidity will be critically reconsidered by taking into account molecular dynamics calculations of heat capacity and conformational entropy, salt bridge networks, electrostatic interactions in folded and unfolded states, and the emerging picture of protein thermostability in view of recently developed network theories. Last, but not least, the influence of high temperature on the active site and activity of enzymes will be considered.

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“…It has been suggested earlier that thermostable proteins show ‘anomalous rigidity’ . This assumption appears to be in strong contrast to the study by Seewald et al .…”

Section: Heat Capacity Configurational Entropy and Fluctuations In Pcontrasting

confidence: 70%

Section: Heat Capacity Configurational Entropy and Fluctuations In Pmentioning

confidence: 99%

Rigidity versus flexibility: the dilemma of understanding protein thermal stability

2015

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“…This may be a reflection of the high thermal stability of the B1 domain and the absence of significant slow timescale motions over the 0-50 8C temperature range. We know of three previous studies focusing on the variation of NH group order parameters with temperature~Man-del et al., 1996;Yang et al, 1997;Evenäs et al, 1999!. For RNase H~Mandel et al, 1996!, the variation was described in terms of a characteristic temperature~T *!…”

Section: Heat Capacities and Characteristic Temperaturesmentioning

confidence: 99%

“…Previously, it has been suggested that thermostable proteins have "anomalous rigidity"~Daniel et al., 1996;Jaenicke, 1996;Cowan, 1997!. This contention appears in stark contrast to the present proposal that~at least in certain cases!…”

Section: Influence Of Heat Capacity On Thermal Stabilitymentioning

confidence: 99%

The role of backbone conformational heat capacity in protein stability: Temperature dependent dynamics of the B1 domain of Streptococcal protein G

et al. 2000

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The contributions of backbone NH group dynamics to the conformational heat capacity of the B1 domain of Streptococcal protein G have been estimated from the temperature dependence of 15 N NMR-derived order parameters. Longitudinal~R 1 ! and transverse~R 2 ! relaxation rates, transverse cross-relaxation rates~h xy !, and steady state $ 1 H%-15 N nuclear Overhauser effects were measured at temperatures of 0, 10, 20, 30, 40, and 50 8C for 89-100% of the backbone secondary amide nitrogen nuclei in the B1 domain. The ratio R 2 0h xy was used to identify nuclei for which conformational exchange makes a significant contribution to R 2 . Relaxation data were fit to the extended model-free dynamics formalism, incorporating an axially symmetric molecular rotational diffusion tensor. The temperature dependence of the order parameter~S 2 ! was used to calculate the contribution of each NH group to conformational heat capacity~C p ! and a characteristic temperature~T *!, representing the density of conformational energy states accessible to each NH group. The heat capacities of the secondary structure regions of the B1 domain are significantly higher than those of comparable regions of other proteins, whereas the heat capacities of less structured regions are similar to those in other proteins. The higher local heat capacities are estimated to contribute up to ;0.8 kJ0mol K to the total heat capacity of the B1 domain, without which the denaturation temperature would be ;9 8C lower~78 8C rather than 87 8C!. Thus, variation of backbone conformational heat capacity of native proteins may be a novel mechanism that contributes to high temperature stabilization of proteins.Keywords: B1 domain; entropy; heat capacity; NMR relaxation; order parameter; protein dynamics; protein stability Most globular proteins are marginally stable because the factors that favor formation of the native state, primarily desolvation of hydrophobic groups and formation of intramolecular hydrogen bonds and salt bridges, are almost equally balanced against those that favor denaturation, primarily the higher conformational entropy of the unfolded protein chain relative to that of the native state~Creigh-ton, 1993; Fersht, 1999!. At high temperatures, the balance between these factors is altered such that many proteins exhibit reversible thermal denaturation with a characteristic midpoint, the melting temperature~T m !. The free energy of unfolding of a proteiñ DG NϪU ! depends upon the changes in enthalpy, entropy, and heat capacity that occur upon unfolding and the temperature~T ! according to the equation~Creighton, 1993; Fersht, 1999!:in which DH 0 and DS 0 are the enthalpy and entropy changes, respectively, at a reference temperature T 0 and DC p,NϪU is the change in heat capacity at constant pressure, which is assumed to be invariant with temperature. Equation 1 indicates that native proteins may be stabilized by an increase in DH 0 or by a decrease in either DS 0 or DC p,NϪU . Thus, one possible strategy for a protein to achieve high thermal ...

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“…Large changes in thermal stability often reflect the accumulation of small contributions made by many mutations. The effects of temperature on the forces contributing to protein stability are many and highly complex [4].…”

Section: Introductionmentioning

confidence: 99%

The effects of multiple ancestral residues on the Thermus thermophilus 3‐isopropylmalate dehydrogenase

Watanabe

Yamagishi

2006

FEBS Letters

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Previously, we showed that mutants of Thermus thermophilus 3-isopropylmalate dehydrogenase (IPMDH) each containing a residue (ancestral residue) that had been predicted to exist in a postulated common ancestor protein often have greater thermal stabilities than does the contemporary wild-type enzyme. In this study, the combined effects of multiple ancestral residues were analyzed. Two mutants, containing multiple mutations, Sup3mut (Val181Thr/Pro324Thr/Ala335Glu) and Sup4mut (Leu134Asn/Val181Thr/Pro324Thr/Ala335Glu) were constructed and show greater thermal stabilities than the wild-type and single-point mutant IPMDHs do. Most of the mutants have similar or improved catalytic efficiencies at 70°C when compared with the wild-type IPMDH.

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Glyceraldehyde-3-phosphate dehydrogenase from Thermotoga maritima: Strategies of protein stabilization

Cited by 9 publications

References 23 publications

Rigidity versus flexibility: the dilemma of understanding protein thermal stability

Rigidity versus flexibility: the dilemma of understanding protein thermal stability

The role of backbone conformational heat capacity in protein stability: Temperature dependent dynamics of the B1 domain of Streptococcal protein G

The effects of multiple ancestral residues on the Thermus thermophilus 3‐isopropylmalate dehydrogenase

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