Engineering thermostability: lessons from thermophilic proteins

Russell, Rosemary; Taylor, G.L.

doi:10.1016/0958-1669(95)80064-6

Cited by 101 publications

(57 citation statements)

References 20 publications

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“…Other factors, such as the number of insertions and deletions in the sequences/ structures, do not appear to be correlated with differences in stability ( Figure 5). AcyMBP is a monomer (confirmed here by dynamic light scattering), and so improved interactions within a multimer 35 are not a viable route to its stabilization.…”

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confidence: 99%

X-ray Structures of the Maltose–Maltodextrin-binding Protein of the Thermoacidophilic Bacterium Alicyclobacillus acidocaldarius Provide Insight into Acid Stability of Proteins

Schäfer

Magnusson

Scheffel

et al. 2004

Journal of Molecular Biology

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Maltose-binding proteins act as primary receptors in bacterial transport and chemotaxis systems. We report here crystal structures of the thermoacidostable maltose-binding protein from Alicyclobacillus acidocaldarius, and explore its modes of binding to maltose and maltotriose. Further, comparison with the structures of related proteins from Escherichia coli (a mesophile), and two hyperthermophiles (Pyrococcus furiosus and Thermococcus litoralis) allows an investigation of the basis of thermo-and acidostability in this family of proteins.The thermoacidophilic protein has fewer charged residues than the other three structures, which is compensated by an increase in the number of polar residues. Although the content of acidic and basic residues is approximately equal, more basic residues are exposed on its surface whereas most acidic residues are buried in the interior. As a consequence, this protein has a highly positive surface charge. Fewer salt bridges are buried than in the other MBP structures, but the number exposed on its surface does not appear to be unusual. These features appear to be correlated with the acidostability of the A. acidocaldarius protein rather than its thermostability.An analysis of cavities within the proteins shows that the extremophile proteins are more closely packed than the mesophilic one. Proline content is slightly higher in the hyperthermophiles and thermoacidophiles than in mesophiles, and this amino acid is more common at the second position of b-turns, properties that are also probably related to thermostability. Secondary structural content does not vary greatly in the different structures, and so is not a contributing factor.

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confidence: 99%

X-ray Structures of the Maltose–Maltodextrin-binding Protein of the Thermoacidophilic Bacterium Alicyclobacillus acidocaldarius Provide Insight into Acid Stability of Proteins

Schäfer

Magnusson

Scheffel

et al. 2004

Journal of Molecular Biology

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“…The rational approach to stabilization is extremely dif®cult, since it is not generally possible to predict the energetic and structural response to mutation in proteins, although the statistics of isolated helices and parts of sheets are predictable to varying degrees (Munoz et al, 1996;Regan et al, 1996). Some insight into protein stabilization has also been gained from the structural comparison of thermophilic proteins with their mesophilic counterparts (Russell & Taylor, 1995). However, the dif®culty underlying this approach is that the sequence divergence between these proteins is prohibitively too large to quantitate the contribution to protein stability on the basis of individual residues.…”

Section: Introductionmentioning

confidence: 99%

Stabilization of GroEL minichaperones by core and surface mutations

Wang

Buckle

Fersht

2000

Journal of Molecular Biology

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We report the crystal structures of two hexa-substituted mutants of a GroEL minichaperone that are more stable than wild-type by 7.0 and 6.1 kcal mol À1 . Their structures imply that the increased stability results from multiple factors including improved hydrophobic packing, optimised hydrogen bonding and favourable structural rearrangements. It is commonly believed that protein core residues are immutable and generally optimized for energy, while on the contrary, surface residues are variable and hence unimportant for stability. But, it is now becoming clear that mutations of both core and surface residues can increase protein stability, and that protein cores are more¯exible and thus more tolerant to mutation than expected. Sequence comparison of homologous proteins has provided a way to pinpoint the residues that contribute constructively to stability and to guide the engineering of protein stability. Stabilizing mutations identi®ed by this approach are most frequently located at protein surfaces but with a few found in protein cores. In the latter case, local¯exibility in the hydrophobic core is the key factor that allows the energetically favourable burial of larger hydrophobic sidechains without undue energetic penalties from steric clashes.

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“…Several investigations have been carried out to illustrate the influence of amino acid on protein thermostability. Russell [1] compared the amino acid composition of citrate synthase from pig, T. acidophilum and P. furiosus. He concluded that as optimal temperature increased, so did the Ile, Tyr, Lys, and Glu content, but the content of thermolabile residues (i.e., Asn, Gln, and Cys) reduced.…”

Section: Introductionmentioning

confidence: 99%

The influence of dipeptide composition on protein thermostability

et al. 2004

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In this work, the influence of dipeptide composition on protein thermostability was studied. After comparing the normalized dipeptide composition between mesophilic proteins and (hyper)thermophilic proteins, we concluded that when organism optimal growth temperature increased, for archaeal proteins, the compositions of VK, KI, YK, IK, KV, KY, and EV increased significantly and the compositions of DA, AD, TD, DD, DT, HD, DH, DR, and DG decreased significantly; and for bacterial proteins, the compositions of KE, EE, EK, YE, VK, KV, KK, LK, EI, EV, RK, EF, KY, VE, KI, KG, EY, FK, KF, FE, KR, VY, MK, WK, and WE increased significantly and the compositions of WQ, AA, QA, MQ, AW, QW, QQ, RQ, QH, HQ, AD, AQ, WL, QL, HA, and DA decreased significantly. So these characteristic dipeptides are correlative to protein thermostability. At the same time, the influence of single amino acid composition on protein thermostability was also studied for comparison. We found that the influence of single amino acid composition could be deduced from the influence of dipeptide composition. So we thought that the influence of dipeptide composition on protein thermostability is larger than the influence of amino acid composition. The characteristic dipeptides not only describe the dipeptides that influence protein thermostability significantly but also show the relationship among significant single amino acids that influence protein thermostability.

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Engineering thermostability: lessons from thermophilic proteins

Cited by 101 publications

References 20 publications

X-ray Structures of the Maltose–Maltodextrin-binding Protein of the Thermoacidophilic Bacterium Alicyclobacillus acidocaldarius Provide Insight into Acid Stability of Proteins

X-ray Structures of the Maltose–Maltodextrin-binding Protein of the Thermoacidophilic Bacterium Alicyclobacillus acidocaldarius Provide Insight into Acid Stability of Proteins

Stabilization of GroEL minichaperones by core and surface mutations

The influence of dipeptide composition on protein thermostability

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