Carbon 13 Nuclear Magnetic Resonance of Pentapeptides of Glycine Containing Central Residues of Aliphatic Amino Acids

Keim, Philip; Vigna, Robert A.; Marshall, Robert C.; Gurd, Frank R. N.

doi:10.1016/s0021-9258(19)43515-1

Cited by 72 publications

(16 citation statements)

References 64 publications

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“…All of the narrow resonances are at the high-field end of the carbonyl region and correspond most closely to the shifts observed in model peptides (Keim et al, 1973a(Keim et al, ,b, 1974. Some residues, e.g., phenylalanine and lysine, show substantial downfield shifts for residues closer to the hydrophobic core;…”

Section: Discussionsupporting

confidence: 77%

Backbone dynamics of a model membrane protein: assignment of the carbonyl carbon carbon-13 NMR resonances in detergent-solubilized M13 coat protein

Henry¹,

Weiner²,

Sykes³

1987

Biochemistry

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The major coat protein of the filamentous bacteriophage M13 is a 50-residue amphiphilic polypeptide which is inserted, as an integral membrane-spanning protein, in the inner membrane of the Escherichia coli host during infection. 13C was incorporated biosynthetically into a total of 23 of the peptide carbonyls using labeled amino acids (alanine, glycine, lysine, phenylalanine, and proline). The structure and dynamics of carbonyl-labeled M13 coat protein were monitored by 13C nuclear magnetic resonance (NMR) spectroscopy. Assignment of many resonances was achieved by using protease digestion, pH titration, or labeling of the peptide bond with both 13C and 15N. The carbonyl region of the natural-abundance 13C NMR spectrum of M13 coat protein in sodium dodecyl sulfate solution shows approximately eight backbone carbonyl resonances with line widths much narrower than the rest. Three of these more mobile residues correspond to assigned peaks (glycine-3, lysine-48, and alanine-49) in the individual amino acid spectra, and another almost certainly arises from glutamic acid-2. A ninth residue, alanine-1, also gives rise to a very narrow carbonyl resonance if the pH is well above or below the pKa of the terminal amino group. These data suggest that only about four residues at either end of the protein experience large-amplitude spatial fluctuations; the rest of the molecule is essentially rigid on the time scale of the overall rotational tumbling of the protein-detergent complex. The relative exposure of different regions of detergent-bound protein was monitored by limited digestion with proteinase K.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionsupporting

confidence: 77%

Backbone dynamics of a model membrane protein: assignment of the carbonyl carbon carbon-13 NMR resonances in detergent-solubilized M13 coat protein

Henry¹,

Weiner²,

Sykes³

1987

Biochemistry

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“…Of particular interest is the resonance located at 17.4 ppm which represents the methyl resonances of alanine residues. The chemical shift position is characteristic of alanine methyl groups in free peptides (Keim et al, 1973). This resonance appears to be the small single resonance observed in Figure 6B on the high-field shoulder of the main aliphatic band of the partially denatured protein, judging from its intensity.…”

Section: Resultsmentioning

confidence: 80%

Carbon-13 nuclear magnetic resonance study of the binding of carbon-13-enriched tetra-L-alanine haptens to Fab' fragments of anti-poly(L-alanine) antibodies

Geller¹,

Wei²,

Shkuda³

et al. 1980

Biochemistry

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“…Tentative assignments of most of the peaks in the 13C NMR spectrum of the alkali-denatured 7S protein (Figure 1) were made by comparing the observed chemical shifts with those of other diamagnetic proteins, peptides, and free amino acids after the effect of amino acid incorporation into peptides on the chemical shift was corrected for (Freedman et al, 1971(Freedman et al, , 1973Christl and Roberts, 1972;Keim et al, 1973aKeim et al, ,b, 1974; Bradbury and Norton, 1973;Quirt et al, 1974;Deslauriers et al, 1975;Van Binst et al, 1975;Oldfield et al, 1975b;Wüthrich and Baumann, 1976;Richarz and Wüthrich, 1978; Howarth and Lilley, 1978;Baianu et al, 1982;Tatham et al, 1985; Kakalis and Baianu, 1989), as well as by taking into account the amino acid analysis data.…”

Section: Resultsmentioning

confidence: 99%

High-resolution carbon-13 nuclear magnetic resonance study of the soybean 7S storage protein fraction in solution

Kakalis¹,

Bâianu²

1990

J. Agric. Food Chem.

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The molecular structure and dynamics of soybean 7S storage proteins were investigated by highresolution 13C NMR at high fields (7.05 and 11.75 T). The spectrum of the alkali-denatured proteins (pH 12.0) is well-resolved, and tentative assignments of 53 peaks are given. At pH 12.0 and 12 mM ionic strength the proteins dissociate into partially unfolded subunits that exhibit fast local motion (10"9-10"10 s), while they maintain a relatively immobile hydrophobic core. The spectra of "native", multisubunit proteins consist of broad peaks as a result of the slower protein tumbling in solution and the expected chemical shift nonequivalence. There is evidence that Phe residues are in the hydrophobic subunit core, whereas Tyr residues are found at the subunits' interface. No major spectral differences were observed between pH 7.6 and 10.3 (at 0.5 M ionic strength, where proteins exist as trimers) or between 13 mM ionic strength and 0.5 M NaCl (at pH 10.2, where a trimer-hexamer interconversion occurs). Potential applications of these results, and of the 13C NMR techniques, in general, include quality control of soy protein isolates in food engineering and the analysis of complex foods that include soy protein ingredients.

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Carbon 13 Nuclear Magnetic Resonance of Pentapeptides of Glycine Containing Central Residues of Aliphatic Amino Acids

Cited by 72 publications

References 64 publications

Backbone dynamics of a model membrane protein: assignment of the carbonyl carbon carbon-13 NMR resonances in detergent-solubilized M13 coat protein

Backbone dynamics of a model membrane protein: assignment of the carbonyl carbon carbon-13 NMR resonances in detergent-solubilized M13 coat protein

Carbon-13 nuclear magnetic resonance study of the binding of carbon-13-enriched tetra-L-alanine haptens to Fab' fragments of anti-poly(L-alanine) antibodies

High-resolution carbon-13 nuclear magnetic resonance study of the soybean 7S storage protein fraction in solution

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