Carbon‐13 nuclear magnetic resonance relaxation study of chymotrypsin inhibitor 2 (CI‐2)

Leatherbarrow, Robin J.; Matthews, Stephen

doi:10.1002/mrc.1260301215

Cited by 4 publications

(4 citation statements)

References 41 publications

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“…The parameters derived from the analysis are given in Table III. The value obtained for the correlation time for overall tumbling (rc = 3.9 ns) is in agreement with literature values for proteins (Richarz et al, 1980;Kay et al, 1989;Clore et al, 1990) and the results of phenylalanine and glycine relaxation studies on CI-2 (Leatherbarrow & Matthews, 1992).…”

Section: "-supporting

confidence: 90%

“…First, hydrogen bond capability is removed by the replacement of the arginine at position 67 by alanine (Figure 1). The resulting reduction in stability and changes to the dynamics of a neighboring phenylalanine ring have already been characterized for this mutant (Jandu et al, 1990; Leatherbarrow & Matthews, 1992). Secondly, modification of the side chain of isoleucine 39 residue within the hydrophobic core of the molecule.…”

mentioning

confidence: 87%

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Carbon-13 NMR study of the effects of mutation on the tryptophan dynamics in chymotrypsin inhibitor 2: correlations with structure and stability

Matthews

Jandu²,

Leatherbarrow³

1993

Biochemistry

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Recombinant chymotrypsin inhibitor 2 (CI-2) and the three mutants Ile39-->Val, Ile39-->Leu, and Arg67-->Ala were successfully enriched with [2-13C]tryptophan at position 24 within the hydrophobic core of the protein. Carbon-13 NMR relaxation measurements were then used to investigate the effect of these mutations on the dynamics of the tryptophan residue. In addition, the stability of wild-type and mutant CI-2s was measured by their susceptibility to unfolding by guanidine hydrochloride. The mutant proteins were all found to be less stable, giving delta delta GU values relative to wild-type of 1.17, 1.96, and 1.21 kcal mol-1, respectively. The indole moiety of the tryptophan residue was found to be more mobile in all the mutants studied than in wild-type CI-2. Order parameters of 0.69, 0.60, 0.56, and 0.44 were derived for wild-type, Ile39-->Val, Ile39-->Leu, and Arg67-->Ala CI-2, respectively. It is concluded that there is a correlation between the protein stability and the picosecond dynamics within the hydrophobic core and that mutations can influence the dynamic behavior of the residues that are relatively distant in the three-dimensional structure.

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Section: "-supporting

confidence: 90%

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confidence: 87%

Carbon-13 NMR study of the effects of mutation on the tryptophan dynamics in chymotrypsin inhibitor 2: correlations with structure and stability

Matthews

Jandu²,

Leatherbarrow³

1993

Biochemistry

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“…Dynamic Perturbation Response. Mutation has long been used as a tool for probing molecular behavior, and this and other studies (20,21,60) extend its use into the realm of dynamics. Recent studies with protein G have demonstrated that a series of surface mutations can be used as a probe of related backbone motions (60).…”

Section: Discussionmentioning

confidence: 99%

“…Findings of this kind are not limited to ligand binding; experiments with protein L have revealed marked changes in methyl dynamics on the nanosecond time scale at sites distal to point mutations (20). Additionally, experiments with chymotrypsin inhibitor 2 revealed changes in the dynamics of a tryptophan side chain in response to an R f A mutation 13 Å away (21). Though these discoveries may seem to be somewhat startling, they have been preceded by findings in molecular dynamics simulations (22)(23)(24).…”

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confidence: 99%

Long-Range Dynamic Effects of Point Mutations Propagate through Side Chains in the Serine Protease Inhibitor Eglin c

2004

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Long-range interactions are fundamental to protein behaviors such as cooperativity and allostery. In an attempt to understand the role protein flexibility plays in such interactions, the distribution of local fluctuations in a globular protein was monitored in response to localized, nonelectrostatic perturbations. Two valine-to-alanine mutations were introduced into the small serine protease inhibitor eglin c, and the (15)N and (2)H NMR spin relaxation properties of these variants were analyzed in terms of the Lipari-Szabo dynamics formalism and compared to those of the wild type. Significant changes in picosecond to nanosecond dynamics were observed in side chains located as much as 13 A from the point of mutation. Additionally, those residues experiencing altered dynamics appear to form contiguous surfaces within the protein. In the case of V54A, the large-to-small mutation results in a rigidification of connected residues, even though this mutation decreases the global stability. These findings suggest that dynamic perturbations arising from single mutations may propagate away from the perturbed site through networks of interacting side chains. That this is observed in eglin c, a classically nonallosteric protein, suggests that such behavior will be observed in many, if not all, globular proteins. Differences in behavior between the two mutants suggest that dynamic responses will be context-dependent.

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Reversible Inhibitors of Serine Proteinases

Wenzel

Tschesche

1995

Peptides

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Carbon‐13 nuclear magnetic resonance relaxation study of chymotrypsin inhibitor 2 (CI‐2)

Abstract: Carbon-13 T, and NOE measurements were used to study the internal motions of the single phenylalanine residue in CI-2 and in a mutant in which an arginine residue adjacent to the Phe had been replaced with an alanine residue.

Cited by 4 publications

References 41 publications

Carbon-13 NMR study of the effects of mutation on the tryptophan dynamics in chymotrypsin inhibitor 2: correlations with structure and stability

Carbon-13 NMR study of the effects of mutation on the tryptophan dynamics in chymotrypsin inhibitor 2: correlations with structure and stability

Long-Range Dynamic Effects of Point Mutations Propagate through Side Chains in the Serine Protease Inhibitor Eglin c

Reversible Inhibitors of Serine Proteinases

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