Molecular self-interactions of ribonuclease A revealed by isothermal titration calorimetry and self-interaction chromatography – Effects of anisotropy of protein surface charges

Tai Chih Kuo, Yu Chiang Huang, Daumantas Matulis, Wen Yih Chen

Research output: Contribution to journalArticle

Abstract

The roles of anisotropy of surface charge in the attraction (or repulsion) between molecules of same identity are probed by using myoglobin (Mb) and ribonuclease A (RNase A) as model proteins. To show the direction and extent of molecular interactions, we estimated second virial coefficients from the heat of protein dilution in isothermal titration calorimetry (ITC) experiments. While our ITC data of Mb were consistent with the study using popular self-interaction chromatography (SIC), those of RNase A in certain pH and salt settings contradicted. The results of size-exclusion chromatography supported the ITC findings. We propose that high anisotropy of surface charge of RNase A causes acute increase of induced attractive self-interaction at elevated ionic strength. Also, due to the use of surface-immobilized protein, the true in-solution behaviors of protein-protein interactions of proteins with high anisotropy of surface charge might not be revealed in SIC.

Original languageEnglish
JournalJournal of the Taiwan Institute of Chemical Engineers
DOIs
Publication statusAccepted/In press - Jan 1 2018

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Pancreatic Ribonuclease
Calorimetry
Surface charge
Chromatography
Titration
Anisotropy
Proteins
Myoglobin
Immobilized Proteins
Molecular interactions
Size exclusion chromatography
Ionic strength
Dilution
Salts
Ribonucleases
Molecules
Experiments

Keywords

  • Charge anisotropy
  • Isothermal titration calorimetry
  • Molecular self-interaction
  • Ribonuclease A
  • Second virial coefficient

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)

Cite this

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title = "Molecular self-interactions of ribonuclease A revealed by isothermal titration calorimetry and self-interaction chromatography – Effects of anisotropy of protein surface charges",
abstract = "The roles of anisotropy of surface charge in the attraction (or repulsion) between molecules of same identity are probed by using myoglobin (Mb) and ribonuclease A (RNase A) as model proteins. To show the direction and extent of molecular interactions, we estimated second virial coefficients from the heat of protein dilution in isothermal titration calorimetry (ITC) experiments. While our ITC data of Mb were consistent with the study using popular self-interaction chromatography (SIC), those of RNase A in certain pH and salt settings contradicted. The results of size-exclusion chromatography supported the ITC findings. We propose that high anisotropy of surface charge of RNase A causes acute increase of induced attractive self-interaction at elevated ionic strength. Also, due to the use of surface-immobilized protein, the true in-solution behaviors of protein-protein interactions of proteins with high anisotropy of surface charge might not be revealed in SIC.",
keywords = "Charge anisotropy, Isothermal titration calorimetry, Molecular self-interaction, Ribonuclease A, Second virial coefficient",
author = "Kuo, {Tai Chih} and Huang, {Yu Chiang} and Daumantas Matulis and Chen, {Wen Yih}",
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AU - Kuo, Tai Chih

AU - Huang, Yu Chiang

AU - Matulis, Daumantas

AU - Chen, Wen Yih

PY - 2018/1/1

Y1 - 2018/1/1

N2 - The roles of anisotropy of surface charge in the attraction (or repulsion) between molecules of same identity are probed by using myoglobin (Mb) and ribonuclease A (RNase A) as model proteins. To show the direction and extent of molecular interactions, we estimated second virial coefficients from the heat of protein dilution in isothermal titration calorimetry (ITC) experiments. While our ITC data of Mb were consistent with the study using popular self-interaction chromatography (SIC), those of RNase A in certain pH and salt settings contradicted. The results of size-exclusion chromatography supported the ITC findings. We propose that high anisotropy of surface charge of RNase A causes acute increase of induced attractive self-interaction at elevated ionic strength. Also, due to the use of surface-immobilized protein, the true in-solution behaviors of protein-protein interactions of proteins with high anisotropy of surface charge might not be revealed in SIC.

AB - The roles of anisotropy of surface charge in the attraction (or repulsion) between molecules of same identity are probed by using myoglobin (Mb) and ribonuclease A (RNase A) as model proteins. To show the direction and extent of molecular interactions, we estimated second virial coefficients from the heat of protein dilution in isothermal titration calorimetry (ITC) experiments. While our ITC data of Mb were consistent with the study using popular self-interaction chromatography (SIC), those of RNase A in certain pH and salt settings contradicted. The results of size-exclusion chromatography supported the ITC findings. We propose that high anisotropy of surface charge of RNase A causes acute increase of induced attractive self-interaction at elevated ionic strength. Also, due to the use of surface-immobilized protein, the true in-solution behaviors of protein-protein interactions of proteins with high anisotropy of surface charge might not be revealed in SIC.

KW - Charge anisotropy

KW - Isothermal titration calorimetry

KW - Molecular self-interaction

KW - Ribonuclease A

KW - Second virial coefficient

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