Raman scattering of native and thermally denatured lysozyme

Brunner, H.; Sussner, H.

doi:10.1016/0005-2795(72)90128-6

Cited by 46 publications

(18 citation statements)

References 13 publications

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“…This peak is consistent with the amino acid residue histidine (Brunner and Sussner, 1972;Lord and Yu, 1970;Tu, 1982). Histidine is catalytically essential in trypsin (Voet et al, 1999) and changes in this site could be reflected by a change in relative activity of the protein.…”

Section: Ft-ramansupporting

confidence: 78%

Influence of protein on mannitol polymorphic form produced during co-spray drying

Hulse

Forbes

Bonner

et al. 2009

International Journal of Pharmaceutics

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Section: Ft-ramansupporting

confidence: 78%

Influence of protein on mannitol polymorphic form produced during co-spray drying

Hulse

Forbes

Bonner

et al. 2009

International Journal of Pharmaceutics

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“…Tropomyosin solution concentrations needed for calculating the molar ellipticity of CD bands were based on an extinction coefficient of 3.1 dl/cm-g a t 278 nm.I9 The Raman spectrometer and sampling techniques have been described elsewhere.la A Spectra-Physics model 165 argon ion laser tuned to 5145. 3 …”

Section: Methodsmentioning

confidence: 99%

“…'- 19 The amide I and I11 regions of the spectrum are potentially useful for conformational analysis. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] Globular proteins contain a mixture of secondary conformations and the assignment of Raman lines to specific secondary conformations such as the alpha-helix, beta-sheet, and disordered conformations has been the aim of these initial studies. Bellocq, Lord, and Mendelsohn4 reported the Raman spectra of bovine serum albumin and beta-lactoglobulin and tentatively assigned specific lines near 1265-1275 cm-I, 1260 cm-', and 1245 cm-I, respectively, to the amide I11 mode of the alpha-helix, beta-sheet, and disordered conformation.…”

Section: Introductionmentioning

confidence: 99%

Raman spectroscopic study of tropomyosin denaturation

Frushour

Koenig

1974

Biopolymers

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SynopsisRaman spectra of the pH denaturation of tropomyosin are presented. In the native state tropomyosin has an alpha-helical content of nearly 90%, but this value drops rapidly as the pH is raised above 9.5. The Raman spectrum of the native state is characterized by a strong amide I line appearing a t 1655 cm-1, very weak scattering in the amide I11 region around 1250 cm-1, and a medium-intensity line a t 940 cm-1. When the protein is pH-denatured, a strong amide I11 line appears a t 1254 cm-l and the 940 cm-l line becomes weak. The intensities of the latter two lines are a sensitive measure of the alpha-helical and disordered chain content. These results are consistent with the helixto-coil studies of the polypeptides.The Raman spectra of a-casein and prothrombin, proteins thought to have little or no ordered secondary structure, are investigated. The amide I11 regions of both spectra display strong lines a t 1254 cm-1 and only weak scattering is observed a t 940 cm-l, features characteristic of the denatured tropomyosin spectrum. The amide I mode of acasein appears a t 1668 cm-1, in agreement with the previously reported spectra of disordered polypeptides, poly-cglutamic acid and poly-Llysine a t pH 7.0 and mechanically deformed poly-Lalanine. 1809

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“…Assignments of S-S and C-S stretching frequencies have been made for biological molecules as complex as lysozyme (8,11), ribonuclease (12, 13), a-chymotrypsin (12), and other proteins (14)(15)(16)(17)(18).…”

mentioning

confidence: 99%

Disulfide Bond Dihedral Angles from Raman Spectroscopy

Wart

Lewis

Scheraga

et al. 1973

Proc. Natl. Acad. Sci. U.S.A.

169

103

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Raman spectra of several compounds containing the CS-SC moiety were obtained (in the solid phase) from 450-800 cm-' to investigate the S-S and C-S stretching behavior. The S-S stretching frequency varied linearly with the CS-SC dihedral angle (obtained from either x-ray or neutron diffraction or ultraviolet absorption) for compounds whose CC-SS dihedral angles were not very different. The ratio of the intensities of the S-S and C-S stretching bands exhibited no recognizable correlation with either the CS-SC dihedral angle or the CSS bond angle, probably because this ratio is sesisitive to the crystalline environment. The linear dependence of the S-S stretching frequency on dihedral angle leads to a dihedral angle for the plant hormone, malformin A, that is in excellent agreement with that estimated from the longest wavelength CS-SC ultraviolet absorption band.The disulfide bond is an important structural feature of many important biological molecules. The CS-SC moiety occurs in antibiotics such as gliotoxin, sporidesmin, and acetylaranotin in the form of an S-S bridged piperazinedione system in which the CS-SC dihedral angle is severely restricted by ring closure (1, 2). Hormones, such as oxytocin (3), vasopressin (4), and malformin (5), contain disulfide bonds (cystine) that can assume conformations with widely varying dihedral angle, depending upon ring size and other structural constraints. In globular proteins, such as ribonuclease, chymotrypsin, lysozyme, carboxypeptidase, etc., packing requirements may result in dihedral angles that vary substantially from the widely accepted value (6) of about 900 for L-cystine in aqueous solution. In chymotrypsin, for example, this angle varies from 87°-126°(7). It is, therefore, of considerable importance to have a method that can provide information about this dihedral angle both in solid state and in aqueous solution. Raman spectroscopy appears to be a suitable technique for this purpose, since the Raman spectra of compounds containing disulfide bonds usually show welldefined bands that arise from C-S and S-S stretching modes (8,9), and these vibrations might be sensitive to the CS-SC dihedral angle.Absorption spectra can also be used to obtain information about CS-SC dihedral angles, since the relationship between this dihedral angle and the longest-wavelength CS-SC absorption frequency in the ultraviolet is well established both theoretically (10) and experimentally (see references cited in ref. 10). However, this method for determining CS-SC dihedral angles is limited to simple systems in which the UV absorption spectrum is free from nondisulfide-related transitions from about 250 to 380 nm. Many systems (proteins for example, which contain aromatic residues) have nondisulfide chromophores that absorb in this region, and thus prevent study of the CS-SC absorption bands. On the other hand, as t To whom requests for reprints should be addressed. 2619has been pointed out (8, 9), these compounds (including proteins) exhibit S-S and C-S stretching bands in a reg...

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Raman scattering of native and thermally denatured lysozyme

Cited by 46 publications

References 13 publications

Influence of protein on mannitol polymorphic form produced during co-spray drying

Influence of protein on mannitol polymorphic form produced during co-spray drying

Raman spectroscopic study of tropomyosin denaturation

Disulfide Bond Dihedral Angles from Raman Spectroscopy

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