Determination of Phosphorylation and Deprotonation Induced Higher Order Structural Transitions in αs‑Caseins

Chemistry

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Rights statement: This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/doi/full/10.1021/acs.analchem.9b03457
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Determination of Phosphorylation and Deprotonation Induced Higher Order Structural Transitions in αs‑Caseins. / Ettah, Ilokugbe; Ashton, Lorna.
In: Analytical Chemistry, Vol. 91, No. 21, 01.11.2019, p. 13940-13946.

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

Bibtex

@article{ff44ec25be7845d8bef4aa13716d998f,

title = "Determination of Phosphorylation and Deprotonation Induced Higher Order Structural Transitions in αs‑Caseins",

abstract = "One extremely sensitive and highly successful application of Raman spectroscopy is the structural characterization of proteins. Understanding higher order structure and its effect on protein stability is essential not only for biopharmaceutical and food manufacturing but also for the understanding of diseases that result from the misfolding of proteins including diabetes type II, Alzheimer's, and Parkinson's disease. Due to the large amount of structural information available in Raman spectra, even small alterations in protein conformations including increased exposure of binding regions or changes in geometry of secondary structural elements can be identified. In this study, we demonstrate the unique sensitivity of Raman spectroscopy to subtle structural transitions in an intrinsically open, flexible protein, α s-casein, in response to phosphorylation and deprotonation. Through the application of 2D correlation analysis two separate transition phases have been identified from pH 6-9 and pH 10-12 for both phosphorylated and dephosphorylated α s-casein. However, the actual structural changes observed in each pH range differed considerably between the phosphorylated and dephosphorylated α s-casein. Furthermore, the presence of the phosphorylated serine residues is demonstrated to have a shielding effect during deprotonation of the protein. ",

author = "Ilokugbe Ettah and Lorna Ashton",

note = "This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright {\textcopyright} American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/doi/full/10.1021/acs.analchem.9b03457 ",

year = "2019",

month = nov,

day = "1",

doi = "10.1021/acs.analchem.9b03457",

language = "English",

volume = "91",

pages = "13940--13946",

journal = "Analytical Chemistry",

issn = "0003-2700",

publisher = "American Chemical Society",

number = "21",

}

RIS

TY - JOUR

T1 - Determination of Phosphorylation and Deprotonation Induced Higher Order Structural Transitions in αs‑Caseins

AU - Ettah, Ilokugbe

AU - Ashton, Lorna

N1 - This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/doi/full/10.1021/acs.analchem.9b03457

PY - 2019/11/1

Y1 - 2019/11/1

N2 - One extremely sensitive and highly successful application of Raman spectroscopy is the structural characterization of proteins. Understanding higher order structure and its effect on protein stability is essential not only for biopharmaceutical and food manufacturing but also for the understanding of diseases that result from the misfolding of proteins including diabetes type II, Alzheimer's, and Parkinson's disease. Due to the large amount of structural information available in Raman spectra, even small alterations in protein conformations including increased exposure of binding regions or changes in geometry of secondary structural elements can be identified. In this study, we demonstrate the unique sensitivity of Raman spectroscopy to subtle structural transitions in an intrinsically open, flexible protein, α s-casein, in response to phosphorylation and deprotonation. Through the application of 2D correlation analysis two separate transition phases have been identified from pH 6-9 and pH 10-12 for both phosphorylated and dephosphorylated α s-casein. However, the actual structural changes observed in each pH range differed considerably between the phosphorylated and dephosphorylated α s-casein. Furthermore, the presence of the phosphorylated serine residues is demonstrated to have a shielding effect during deprotonation of the protein.

AB - One extremely sensitive and highly successful application of Raman spectroscopy is the structural characterization of proteins. Understanding higher order structure and its effect on protein stability is essential not only for biopharmaceutical and food manufacturing but also for the understanding of diseases that result from the misfolding of proteins including diabetes type II, Alzheimer's, and Parkinson's disease. Due to the large amount of structural information available in Raman spectra, even small alterations in protein conformations including increased exposure of binding regions or changes in geometry of secondary structural elements can be identified. In this study, we demonstrate the unique sensitivity of Raman spectroscopy to subtle structural transitions in an intrinsically open, flexible protein, α s-casein, in response to phosphorylation and deprotonation. Through the application of 2D correlation analysis two separate transition phases have been identified from pH 6-9 and pH 10-12 for both phosphorylated and dephosphorylated α s-casein. However, the actual structural changes observed in each pH range differed considerably between the phosphorylated and dephosphorylated α s-casein. Furthermore, the presence of the phosphorylated serine residues is demonstrated to have a shielding effect during deprotonation of the protein.

U2 - 10.1021/acs.analchem.9b03457

DO - 10.1021/acs.analchem.9b03457

M3 - Journal article

VL - 91

SP - 13940

EP - 13946

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 21

ER -

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