Isotopic exchange probed through NMR spectroscopy has been a precious source of information regarding local protein unfolding processes due to its ability to observe the exchange rate of single residues' amide sites. BLUU-Tramp extends this information allowing to follow, through two series of experiments, how the exchange constants vary over temperature. In this thesis a novel approach is introduced by means of an ad hoc calculation routine which is able to extract thermodynamic parameters from the decays of the amide signals, obtained through suitable treatment of the experimental data by another original routine (TinT) of the suite devised in our laboratory. By assuming that the isotopic exchange rates can be approximated with the EX2 limit, it is possible to estimate both the ΔG of local unfolding and other underlying thermodynamic parameters, in particular the ΔCp. The method is used to obtain these parameters in two human proteins, β2-microglobulin and lysozyme. Furthermore, a similar computational approach is presented which is able to extract thermodynamic parameters from classical isotopic exchange experiments carried out at various different temperatures. Comparisons among the results extracted from BLUU-Tramp series and classical exchange experiments are drawn and critical aspects in the latter kind of experiments are discussed

BLUU-Tramp data analysis: thermodynamic landscape of local unfolding processes / Marco Vuano - Udine. , 2016 Apr 01. 28. ciclo

BLUU-Tramp data analysis: thermodynamic landscape of local unfolding processes

Vuano, Marco
2016-04-01

Abstract

Isotopic exchange probed through NMR spectroscopy has been a precious source of information regarding local protein unfolding processes due to its ability to observe the exchange rate of single residues' amide sites. BLUU-Tramp extends this information allowing to follow, through two series of experiments, how the exchange constants vary over temperature. In this thesis a novel approach is introduced by means of an ad hoc calculation routine which is able to extract thermodynamic parameters from the decays of the amide signals, obtained through suitable treatment of the experimental data by another original routine (TinT) of the suite devised in our laboratory. By assuming that the isotopic exchange rates can be approximated with the EX2 limit, it is possible to estimate both the ΔG of local unfolding and other underlying thermodynamic parameters, in particular the ΔCp. The method is used to obtain these parameters in two human proteins, β2-microglobulin and lysozyme. Furthermore, a similar computational approach is presented which is able to extract thermodynamic parameters from classical isotopic exchange experiments carried out at various different temperatures. Comparisons among the results extracted from BLUU-Tramp series and classical exchange experiments are drawn and critical aspects in the latter kind of experiments are discussed
1-apr-2016
NMR; Isotopic; Exchange; BLUU-Tramp: Data Analysis
BLUU-Tramp data analysis: thermodynamic landscape of local unfolding processes / Marco Vuano - Udine. , 2016 Apr 01. 28. ciclo
File in questo prodotto:
File Dimensione Formato  
10990_680_Tesi_Marco_Vuano_Corretta.pdf

accesso aperto

Tipologia: Tesi di dottorato
Licenza: Non specificato
Dimensione 15.52 MB
Formato Adobe PDF
15.52 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11390/1132896
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? ND
social impact