Reassignment of the histidine 1H nuclear magnetic resonances of ribonuclease-A

Bradbury, J. H.; Teh, Joo Seng

doi:10.1039/c39750000936

Cited by 10 publications

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“…see Fig. 8) and the S-protein produced from it gave an area for this resonance that agrees with this assignment [7]. The C-2 protons of residues 105 and 119 of RNase-A after partial deuteration were 28% and 47% respectively of their original size.…”

Section: Studies On S-protein From Rnase-asupporting

confidence: 70%

“…2, 4 and 6). The confirmation of this assignment comes from the partial deuteration experiments described previously [7] in which RNase-A was deuterated at pH 7.15 for 6 days at 35 "C. Histidine-48 was the only histidine residue not deuterated to any observable degree (in agreement with the rate data. see Fig.…”

Section: Studies On S-protein From Rnase-asupporting

confidence: 69%

“…The amount of hydrogen-deuterium exchange increases with increasing times of incubation at 35 "C but this effect cannot be quantified because of the occurrence of a new peak X that also increases with time. This peak is either absent or present to only a very small degrce in native RNase-A [7] (and Fig. 1 of [16]) and has also been observed on acid denaturation at pH < 3 [32] or on reversible heat denaturation at pH 3 [33].…”

Section: Stuhility Of Rnuse To Alkaline Solutionmentioning

confidence: 99%

“…The assignment of the C-2 histidine resonances in S-protein has been indicated very briefly [7] and will now be considered in detail. The upfield broad resonance is readily assigned to histidine-48 because of its similar position and breadth to histidine-48 in RNase-A (see Fig.…”

Section: Studies On S-protein From Rnase-amentioning

confidence: 99%

“…We have therefore studied the titration of the histidine residues of S-peptide and S-protein by ' H NMR spectroscopy using some samples partially dcuterated at the C-2 position. The latter were originally prepared to check the assignment of the C-2 histidine proton resonances in RNase-A [7] and have also proved useful in the assignment of these resonances in S-protein. A recent independent study of this subject has appeared [25], but in comparing our data with theirs, it is suggested that their sample of S-protein contains an impurity of irreversibly denatured material.…”

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confidence: 99%

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Nuclear‐Magnetic‐Resonance Study of the Histidine Residues of S‐Peptide and S‐Protein and Kinetics of ¹H‐²H Exchange of Ribonuclease A

Bradbury

Crompton

Teh

1977

European Journal of Biochemistry

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1H NMR spectroscopy at 100 MHz was used to determine the first‐order rate constants for the 1H‐2H exchange of the H‐2 histidine resonances of RNase‐A in 2H2O at 35°C and pH meter readings of 7, 9, 10 and 10.5. Prolonged exposure in 2H2O at 35°C and pH meter reading 11 caused irreversible denaturation of RNase‐A. The rate constants at pH 7 and 9 agreed reasonably well with those obtained in 1H‐3H exchange experiments by Ohe, J., Matsuo, H., Sakiyama, F. and Narita, K. [J. Biochem. (Tokyo) 75, 1197–1200 1974)]. The rate data obtained by various authors is summarised and the reasons for the poor agreement between the data is discussed. The first‐order rate constant for the exchange of His‐48 increases rapidly from near zero at pH 9 (due to its inaccessibility to solvent) with increase of pH to 10.5. The corresponding values for His‐119 show a decrease and those for His‐12 a small increase over the same pH range. These changes are attributed to a conformational change in the hinge region of RNase‐A (probably due to the titration of Tyr‐25) which allows His‐48 to become accessible to solvent. 1H NMR spectra of S‐protein and S‐peptide, and of material partially deuterated at the C‐2 positions of the histidine residues confirm the reassignment of the histidine resonances of RNase‐A [Bradbury, J. H. & Teh, J. S. (1975) Chem. Commun., 936–937]. The chemical shifts of the C‐2 and C‐4 protons of histidine‐12 of S‐peptide are followed as a function of pH and a pK′ value of 6.75 is obtained. The reassignment of the three C‐2 histidine resonances of S‐protein is confirmed by partial deuteration studies. The pK′ values obtained from titration of the H‐2 resonances of His‐48, His‐105 and His‐119 are 5.3, 6.5 and 6.0, respectively. The S‐protein is less stable to acid than RNase‐A since the former, but not the latter, shows evidence of reversible denaturation at pH 3 and 26°C. His‐48 in S‐protein titrates normally and has a lower pK than in RNase‐A probably because of the absence of Asp‐14, which in RNase‐A forms a hydrogen bond with His‐48 and causes it to be inaccessible to solvent, at pH values below 9.

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Section: Studies On S-protein From Rnase-asupporting

confidence: 70%

Section: Studies On S-protein From Rnase-asupporting

confidence: 69%

Section: Stuhility Of Rnuse To Alkaline Solutionmentioning

confidence: 99%

Section: Studies On S-protein From Rnase-amentioning

confidence: 99%

mentioning

confidence: 99%

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Nuclear‐Magnetic‐Resonance Study of the Histidine Residues of S‐Peptide and S‐Protein and Kinetics of ¹H‐²H Exchange of Ribonuclease A

Bradbury

Crompton

Teh

1977

European Journal of Biochemistry

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Kinetics of hydrogen-deuterium exchange of tryptophan and tryptophan peptides in deutero-trifluoroacetic acid using proton magnetic resonance spectroscopy

Norton

Bradbury

1976

Mol Cell Biochem

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The methods which have been used for the observation and assignment of resonances in the NMR spectra of proteins are reviewed. One such method, the selective deuteration of the aromatic protons of tryptophyl residues, is studied by NMR spectroscopy in model compounds in this paper, and in proteins in the following paper. On the basis of a reassignment of the PMR spectrum of the aromatic protons of L-tryptophan, the relative rates of H-D exchange in deutero-trifluoracetic acid (d-TFA) are H-2 greater than H-5 greater than H-6 greater than H-4 approximately H-7. The energies of activation for the first order exchange of both the H-2 and H-5 protons is 12 k.cal.mol-1. The rate constant for exchange of the H-2 protons of tryptophyl residues in peptides is much greater than in the amino acid itself and 5-10 times that for exchange of the H-5 protons. This suggests that the method can be used to label tryptophyl residues in proteins rapidly and specifically.

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1H NMR study on the interaction of cupric ion with ribonuclease A

Sportelli

Viti

1979

Biochimica et Biophysica Acta (BBA) - Protein Structure

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Reassignment of the histidine 1H nuclear magnetic resonances of ribonuclease-A

Cited by 10 publications

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Nuclear‐Magnetic‐Resonance Study of the Histidine Residues of S‐Peptide and S‐Protein and Kinetics of ¹H‐²H Exchange of Ribonuclease A

Nuclear‐Magnetic‐Resonance Study of the Histidine Residues of S‐Peptide and S‐Protein and Kinetics of ¹H‐²H Exchange of Ribonuclease A

Kinetics of hydrogen-deuterium exchange of tryptophan and tryptophan peptides in deutero-trifluoroacetic acid using proton magnetic resonance spectroscopy

1H NMR study on the interaction of cupric ion with ribonuclease A

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Reassignment of the histidine 1H nuclear magnetic resonances of ribonuclease-A

Cited by 10 publications

References 0 publications

Nuclear‐Magnetic‐Resonance Study of the Histidine Residues of S‐Peptide and S‐Protein and Kinetics of 1H‐2H Exchange of Ribonuclease A

Nuclear‐Magnetic‐Resonance Study of the Histidine Residues of S‐Peptide and S‐Protein and Kinetics of 1H‐2H Exchange of Ribonuclease A

Kinetics of hydrogen-deuterium exchange of tryptophan and tryptophan peptides in deutero-trifluoroacetic acid using proton magnetic resonance spectroscopy

1H NMR study on the interaction of cupric ion with ribonuclease A

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Nuclear‐Magnetic‐Resonance Study of the Histidine Residues of S‐Peptide and S‐Protein and Kinetics of ¹H‐²H Exchange of Ribonuclease A

Nuclear‐Magnetic‐Resonance Study of the Histidine Residues of S‐Peptide and S‐Protein and Kinetics of ¹H‐²H Exchange of Ribonuclease A