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The importance of protonation in the investigation of protein phosphorylation using Raman spectroscopy and Raman optical activity

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The importance of protonation in the investigation of protein phosphorylation using Raman spectroscopy and Raman optical activity. / Ashton, Lorna; Johannessen, Christian; Goodacre, Royston.
In: Analytical Chemistry, Vol. 83, No. 20, 15.10.2011, p. 7978-7983.

Research output: Contribution to Journal/Magazine › Journal article › peer-review

Harvard

Ashton, L, Johannessen, C & Goodacre, R 2011, 'The importance of protonation in the investigation of protein phosphorylation using Raman spectroscopy and Raman optical activity', Analytical Chemistry, vol. 83, no. 20, pp. 7978-7983. https://doi.org/10.1021/ac202041f

APA

Ashton, L., Johannessen, C., & Goodacre, R. (2011). The importance of protonation in the investigation of protein phosphorylation using Raman spectroscopy and Raman optical activity. Analytical Chemistry, 83(20), 7978-7983. https://doi.org/10.1021/ac202041f

Vancouver

Ashton L, Johannessen C, Goodacre R. The importance of protonation in the investigation of protein phosphorylation using Raman spectroscopy and Raman optical activity. Analytical Chemistry. 2011 Oct 15;83(20):7978-7983. doi: 10.1021/ac202041f

Author

Ashton, Lorna ; Johannessen, Christian ; Goodacre, Royston. / The importance of protonation in the investigation of protein phosphorylation using Raman spectroscopy and Raman optical activity. In: Analytical Chemistry. 2011 ; Vol. 83, No. 20. pp. 7978-7983.

Bibtex

@article{594e431776d54cb8a3c0c23b878a8f72,
title = "The importance of protonation in the investigation of protein phosphorylation using Raman spectroscopy and Raman optical activity",
abstract = "The effect of protonation on amino acid monomers and protein phosphorylation was studied by means of a combination of Raman scattering and Raman optical activity (ROA). In the past, identifying spectral variations in phosphorylated proteins arising from either the phosphate stretch or amide vibrational modes has proven to be challenging mainly due to the loss of amide and P = O band intensity in the presence of phosphate. By contrast, we have developed a novel strategy based on the careful monitoring of the sample pH and thereby modified the protonation state, such that these difficulties can be overcome and phosphate-derived vibrations are readily visualized with both Raman and ROA. Variations in pH-dependent spectral sets of phosphorylated amino acid monomers serine and threonine demonstrated that the protonation state could be determined by the intensity of the monobasic (-OPO(3)H(-)) phosphate stretch band occurring at similar to 1080 cm(-1) the dibasic (-OPO(3)(2-)) band measured at similar to 980 cm(-1) in both Raman and ROA. Furthermore, by adjustment of the pH of aqueous samples of the phosphoprotein alpha-casein and comparing this result with dephosphorylated alpha-casein, spectral variations in phosphate stretch bands and amide bands could be easily determined. Consequently, structural variations due to both protonation and dephosphorylation could be distinguished, demonstrating the potential of Raman and ROA for future investigations of phosphoprotein structure and interactions.",
keywords = "CASEIN MICELLE STRUCTURE, UNFOLDED PROTEINS, PEPTIDES",
author = "Lorna Ashton and Christian Johannessen and Royston Goodacre",
year = "2011",
month = oct,
day = "15",
doi = "10.1021/ac202041f",
language = "English",
volume = "83",
pages = "7978--7983",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "20",

}

RIS

TY - JOUR

T1 - The importance of protonation in the investigation of protein phosphorylation using Raman spectroscopy and Raman optical activity

AU - Ashton, Lorna

AU - Johannessen, Christian

AU - Goodacre, Royston

PY - 2011/10/15

Y1 - 2011/10/15

N2 - The effect of protonation on amino acid monomers and protein phosphorylation was studied by means of a combination of Raman scattering and Raman optical activity (ROA). In the past, identifying spectral variations in phosphorylated proteins arising from either the phosphate stretch or amide vibrational modes has proven to be challenging mainly due to the loss of amide and P = O band intensity in the presence of phosphate. By contrast, we have developed a novel strategy based on the careful monitoring of the sample pH and thereby modified the protonation state, such that these difficulties can be overcome and phosphate-derived vibrations are readily visualized with both Raman and ROA. Variations in pH-dependent spectral sets of phosphorylated amino acid monomers serine and threonine demonstrated that the protonation state could be determined by the intensity of the monobasic (-OPO(3)H(-)) phosphate stretch band occurring at similar to 1080 cm(-1) the dibasic (-OPO(3)(2-)) band measured at similar to 980 cm(-1) in both Raman and ROA. Furthermore, by adjustment of the pH of aqueous samples of the phosphoprotein alpha-casein and comparing this result with dephosphorylated alpha-casein, spectral variations in phosphate stretch bands and amide bands could be easily determined. Consequently, structural variations due to both protonation and dephosphorylation could be distinguished, demonstrating the potential of Raman and ROA for future investigations of phosphoprotein structure and interactions.

AB - The effect of protonation on amino acid monomers and protein phosphorylation was studied by means of a combination of Raman scattering and Raman optical activity (ROA). In the past, identifying spectral variations in phosphorylated proteins arising from either the phosphate stretch or amide vibrational modes has proven to be challenging mainly due to the loss of amide and P = O band intensity in the presence of phosphate. By contrast, we have developed a novel strategy based on the careful monitoring of the sample pH and thereby modified the protonation state, such that these difficulties can be overcome and phosphate-derived vibrations are readily visualized with both Raman and ROA. Variations in pH-dependent spectral sets of phosphorylated amino acid monomers serine and threonine demonstrated that the protonation state could be determined by the intensity of the monobasic (-OPO(3)H(-)) phosphate stretch band occurring at similar to 1080 cm(-1) the dibasic (-OPO(3)(2-)) band measured at similar to 980 cm(-1) in both Raman and ROA. Furthermore, by adjustment of the pH of aqueous samples of the phosphoprotein alpha-casein and comparing this result with dephosphorylated alpha-casein, spectral variations in phosphate stretch bands and amide bands could be easily determined. Consequently, structural variations due to both protonation and dephosphorylation could be distinguished, demonstrating the potential of Raman and ROA for future investigations of phosphoprotein structure and interactions.

KW - CASEIN MICELLE STRUCTURE

KW - UNFOLDED PROTEINS

KW - PEPTIDES

U2 - 10.1021/ac202041f

DO - 10.1021/ac202041f

M3 - Journal article

VL - 83

SP - 7978

EP - 7983

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 20

ER -