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Title: Characterization of pH titration shifts for all the nonlabile proton resonances in a protein by two-dimensional NMR: The case of mouse epidermal growth factor

Abstract

The pH titration shifts for all the nonlabile proton resonances in a 53-residue protein (mouse epidermal growth factor) were measured in the p{sup 2}H range 1.5-9 with two-dimensional (2D) {sup 1}H NMR. The 2D NMR pH titration experiment made it possible to determine the pK values for all the ionizable group which were titrated in the pH range 1.5-9 in the protein. The pK values of the nine ionizable groups ({alpha}-amino group, four Asp, two Glu, one His, and {alpha}-carboxyl group) were found to be near their normal values. The 2D titration experiment also provided a detailed description of the pH-dependent behavior of the proton chemical shifts and enabled us to characterize the pH-dependent changes of protein conformation. Analysis of the pH-dependent shifts of ca. 200 proton resonances offered evidence of conformational changes in slightly basic pH solution: The deprotonation of the N-terminal {alpha}-amino group induced a widespread conformational change over the {beta}-sheet structure in the protein, while the effects of deprotonation of the His22 imidazole group were relatively localized. The authors found that the 2D NMR pH titration experiment is a powerful tool for investigating the structural and dynamic properties of proteins.

Authors:
; ;  [1]
  1. Tokyo Metropolitan Inst. of Medical Science (Japan)
Publication Date:
OSTI Identifier:
5014451
Resource Type:
Journal Article
Journal Name:
Biochemistry; (United States)
Additional Journal Information:
Journal Volume: 30:20; Journal ID: ISSN 0006-2960
Country of Publication:
United States
Language:
English
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; GROWTH FACTORS; NUCLEAR MAGNETIC RESONANCE; PEPTIDE HORMONES; MOLECULAR STRUCTURE; CHEMICAL SHIFT; EPIDERMIS; IMIDAZOLES; IONIZATION; MICE; PH VALUE; PROTONS; TITRATION; ANIMAL TISSUES; ANIMALS; AZOLES; BARYONS; BODY; ELEMENTARY PARTICLES; EPITHELIUM; FERMIONS; HADRONS; HETEROCYCLIC COMPOUNDS; HORMONES; MAGNETIC RESONANCE; MAMMALS; MITOGENS; NUCLEONS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANS; PROTEINS; RESONANCE; RODENTS; SKIN; TISSUES; VERTEBRATES; 550601* - Medicine- Unsealed Radionuclides in Diagnostics

Citation Formats

Kohda, Daisuke, Sawada, Toshie, and Inagaki, Fuyuhiko. Characterization of pH titration shifts for all the nonlabile proton resonances in a protein by two-dimensional NMR: The case of mouse epidermal growth factor. United States: N. p., 1991. Web. doi:10.1021/bi00234a009.
Kohda, Daisuke, Sawada, Toshie, & Inagaki, Fuyuhiko. Characterization of pH titration shifts for all the nonlabile proton resonances in a protein by two-dimensional NMR: The case of mouse epidermal growth factor. United States. https://doi.org/10.1021/bi00234a009
Kohda, Daisuke, Sawada, Toshie, and Inagaki, Fuyuhiko. 1991. "Characterization of pH titration shifts for all the nonlabile proton resonances in a protein by two-dimensional NMR: The case of mouse epidermal growth factor". United States. https://doi.org/10.1021/bi00234a009.
@article{osti_5014451,
title = {Characterization of pH titration shifts for all the nonlabile proton resonances in a protein by two-dimensional NMR: The case of mouse epidermal growth factor},
author = {Kohda, Daisuke and Sawada, Toshie and Inagaki, Fuyuhiko},
abstractNote = {The pH titration shifts for all the nonlabile proton resonances in a 53-residue protein (mouse epidermal growth factor) were measured in the p{sup 2}H range 1.5-9 with two-dimensional (2D) {sup 1}H NMR. The 2D NMR pH titration experiment made it possible to determine the pK values for all the ionizable group which were titrated in the pH range 1.5-9 in the protein. The pK values of the nine ionizable groups ({alpha}-amino group, four Asp, two Glu, one His, and {alpha}-carboxyl group) were found to be near their normal values. The 2D titration experiment also provided a detailed description of the pH-dependent behavior of the proton chemical shifts and enabled us to characterize the pH-dependent changes of protein conformation. Analysis of the pH-dependent shifts of ca. 200 proton resonances offered evidence of conformational changes in slightly basic pH solution: The deprotonation of the N-terminal {alpha}-amino group induced a widespread conformational change over the {beta}-sheet structure in the protein, while the effects of deprotonation of the His22 imidazole group were relatively localized. The authors found that the 2D NMR pH titration experiment is a powerful tool for investigating the structural and dynamic properties of proteins.},
doi = {10.1021/bi00234a009},
url = {https://www.osti.gov/biblio/5014451}, journal = {Biochemistry; (United States)},
issn = {0006-2960},
number = ,
volume = 30:20,
place = {United States},
year = {Tue May 21 00:00:00 EDT 1991},
month = {Tue May 21 00:00:00 EDT 1991}
}