Assignment of the histidine peaks in the nuclear magnetic resonance spectrum of ribonuclease.

Meadows, Donella H.; Jardetzky, Oleg; Epand, Richard M.; Rüterjans, H.; Scheraga, Harold A.

doi:10.1073/pnas.60.3.766

Cited by 143 publications

(52 citation statements)

References 7 publications

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“…In agreement with other workers [ 11,18,32], resonance H 4 belonging to the C-2 proton of His-48 was observed at neutral pH in RNase S (Fig. 7) but not in RNase A (Fig.…”

Section: Comparison With Rnuse S : Assignments Of H 3 ' Y 3 and F4supporting

confidence: 80%

“…Spectra at lower pH values indicated an apparent pK value between 4 and 5. Probably, resonance I is identical to the peak Meadows et al [11,12] reported to shift upfield on addition of inhibitors. We can rule out its assignment to Tyr-25 1191 since the Y 3 doublets remained at nearly the same positions.…”

Section: Efsects Of Active-site Inhibitors: Assignment O F F 3 ; Resosupporting

confidence: 54%

“…From the phenylalanines in RNase, Phe-120 is most directly involved in the binding of active-site inhibitors 11,34,35]. Therefore, we tentatively assign the phenylalanine resonance that is most strongly affected by inhibitors, F 3, to Phe-120.…”

Section: Efsects Of Active-site Inhibitors: Assignment O F F 3 ; Resomentioning

confidence: 99%

“…Another singlet, H4, which is not observed between pH 5.2 and pH 7.9 (cf. [11,18,19]) belongs to the C-2 proton of His-48 [6,8, 101. Tyrosine rings that flip rapidly around their CS -Cy bonds are represented by two mutually coupled doublets, both corresponding to two protons [30]. Double resonance experiments (Fig.…”

Section: The Aromatic Resonancesmentioning

confidence: 99%

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The Aromatic Residues of Bovine Pancreatic Ribonuclease Studied by ¹H Nuclear Magnetic Resonance

Lenstra

Bolscher

Stob

et al. 1979

European Journal of Biochemistry

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1. The aromatic proton resonances in the 360-MHz 'H nuclear magnetic resonance (NMR) spectrum of bovine pancreatic ribonuclease were divided into histidine, tyrosine and phenylalanine resonances by means of pH titrations and double resonance experiments.2. Photochemically induced dynamic nuclear polarization spectra showed that one histidine (His-1 19) and two tyrosines are accessible to photo-excited flavin. This permitted the identification of the C-4 proton resonance of His-119.3. The resonances of the ring protons of Tyr-25, Tyr-76 and Tyr-1 15 and the C-4 proton of His-12 were identified by comparison with subtilisin-modified and nitrated ribonucleases. Other resonances were assigned tentatively to 4. On addition of active-site inhibitors, all phenylalanine resonances broadened or disappeared. The resonance that was most affected was assigned tentatively to Phe-120.5. Four of the six tyrosines of bovine RNase, identified as Tyr-76, Tyr-115 and, tentatively, Tyr-73 and Tyr-92, are titratable above pH 9. The rings of Tyr-73 and Tyr-115 are rapidly rotating or flipping by 180" about their CP-Cc" bond and are accessible to flavin in photochemically induced dynamic nuclear polarization experiments. Tyr-25 is involved in a pH-dependent conformational transition, together with Asp-14 and His-48. A scheme for this transition is proposed.6. Binding of active-site inhibitors to bovine RNase only influences the active site and its immediate surroundings. These conformational changes are probably not connected with the pH-dependent transition in the region of 7. In NMR spectra of RNase A at elevated temperatures, no local unfolding below the temperature of the thermal denaturation was observed. NMR spectra of thermally unfolded RNase A indicated that the deviations from a random coil are small and might be caused by interactions between neighbouring residues.

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“…In agreement with other workers [ 11,18,32], resonance H 4 belonging to the C-2 proton of His-48 was observed at neutral pH in RNase S (Fig. 7) but not in RNase A (Fig.…”

Section: Comparison With Rnuse S : Assignments Of H 3 ' Y 3 and F4supporting

confidence: 80%

Section: Efsects Of Active-site Inhibitors: Assignment O F F 3 ; Resosupporting

confidence: 54%

Section: Efsects Of Active-site Inhibitors: Assignment O F F 3 ; Resomentioning

confidence: 99%

Section: The Aromatic Resonancesmentioning

confidence: 99%

See 2 more Smart Citations

The Aromatic Residues of Bovine Pancreatic Ribonuclease Studied by ¹H Nuclear Magnetic Resonance

Lenstra

Bolscher

Stob

et al. 1979

European Journal of Biochemistry

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“…Determinations of the apparent dissociation constants K' of the carboxyl groups were made from a plot of log [(S,-S)/(S-S,)] against pH as described previously [13,24], where a , , 6, and 6 are the observed chemical shifts in ppm in the acid and basic forms and at the particular pH of the measurement respectively. The pK' is the value of the pH at which the log term is zero.…”

Section: Determination Of Dissociation Constantsmentioning

confidence: 99%

Introduction of a Strong Binding Site for Lanthanides at the N-Terminus of Peptides and Ribonuclease A

Bradbury¹,

Johnson²,

Warren³

1978

Eur J Biochem

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A new reagent, phthalyl-4-isothiocyanate, reacts specifically at the N-terminus of a peptide or protein with the production of a phthalic acid 4-thiocarbamyl derivative. The adjacent carboxyl groups of the phthalyl group serve as a strong binding site for lanthanides at pH 5 with a stability constant for praseodymium about 220 times that of a single carboxyl group.Reaction of phthalyl-4-isothiocyanate with a-amino groups of peptides at 25 "C in 2H,O at pH meter readings of 8.7 and 7.2 is complete in 15 min and 24 h respectively, without modification of histidine or tyrosine side chains. At the lower pH there is no reaction of the &-amino group of lysine. The phthalic acid 4-thiocarbamyl derivative of ribonuclease A is cyclised and cleaved by the normal Edman degradation in acid to produce the phthalic acid 4-thiohydantoin of lysine. The a-CH proton of lysine-1 in ribonuclease A produces a triplet nuclear magnetic resonance that has been assigned on the basis of its coupling constant, the pH dependence of its chemical shift (pK' is 7.6) and its disappearance on reaction of ribonuclease A with phthalyl-4-isothiocyanate.The reaction of phthalyl-4-isothiocyanate with a protein allows the introduction of a strong lanthanide binding site specifically at the N-terminus, as a starting point for structural studies using nuclear magnetic resonance spectroscopy.

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Biological Macromolecules

Jardetzky¹

2007

Encyclopedia of Magnetic Resonance

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Assignment of the histidine peaks in the nuclear magnetic resonance spectrum of ribonuclease.

Cited by 143 publications

References 7 publications

The Aromatic Residues of Bovine Pancreatic Ribonuclease Studied by ¹H Nuclear Magnetic Resonance

The Aromatic Residues of Bovine Pancreatic Ribonuclease Studied by ¹H Nuclear Magnetic Resonance

Introduction of a Strong Binding Site for Lanthanides at the N-Terminus of Peptides and Ribonuclease A

Biological Macromolecules

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Assignment of the histidine peaks in the nuclear magnetic resonance spectrum of ribonuclease.

Cited by 143 publications

References 7 publications

The Aromatic Residues of Bovine Pancreatic Ribonuclease Studied by 1H Nuclear Magnetic Resonance

The Aromatic Residues of Bovine Pancreatic Ribonuclease Studied by 1H Nuclear Magnetic Resonance

Introduction of a Strong Binding Site for Lanthanides at the N-Terminus of Peptides and Ribonuclease A

Biological Macromolecules

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The Aromatic Residues of Bovine Pancreatic Ribonuclease Studied by ¹H Nuclear Magnetic Resonance

The Aromatic Residues of Bovine Pancreatic Ribonuclease Studied by ¹H Nuclear Magnetic Resonance