東華大學圖書館 |

語系: 繁體中文

說明(常見問題)

回圖書館首頁

手機版館藏查詢

登入

回首頁

切換: 標籤 | MARC模式 | ISBD

Novel Mechanistic Insights Into the ...

Vasilchuk, Daniel.

FindBook

Google Book

Amazon

博客來

Novel Mechanistic Insights Into the Pressure and Temperature Stabilization of Enzymes.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Novel Mechanistic Insights Into the Pressure and Temperature Stabilization of Enzymes./
作者:	Vasilchuk, Daniel.
面頁冊數:	108 p.
附註:	Source: Dissertation Abstracts International, Volume: 78-02(E), Section: B.
Contained By:	Dissertation Abstracts International78-02B(E).
標題:	Biochemistry. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10158621
ISBN:	9781369136296

Novel Mechanistic Insights Into the Pressure and Temperature Stabilization of Enzymes.
Vasilchuk, Daniel.

Novel Mechanistic Insights Into the Pressure and Temperature Stabilization of Enzymes. - 108 p.

Source: Dissertation Abstracts International, Volume: 78-02(E), Section: B.

Thesis (Ph.D.)--Rensselaer Polytechnic Institute, 2016.

Pressure and temperature have important roles in the evolution of all organisms and their associated proteins. Both factors need to be considered when attempting to engineer more stable proteins for industrial and pharmaceutical applications. In this work, the thermodynamic properties of ancestral and modern proteins were extensively characterized to determine whether the ancestral proteins had unique, or in some way improved, parameters when compared to their modern homologues. The experimental findings, with a primary focus on the pressure-related (volumetric) parameters, could provide clues for engineering more stable proteins.

ISBN: 9781369136296Subjects--Topical Terms:

518028
Biochemistry.

Novel Mechanistic Insights Into the Pressure and Temperature Stabilization of Enzymes.
LDR:04550nmm a2200325 4500 001 2076029
005 20161101084220.5
008 170521s2016 ||||||||||||||||| ||eng d
020 $a 9781369136296
035 $a (MiAaPQ)AAI10158621
035 $a AAI10158621
040 $a MiAaPQ $c MiAaPQ
100 1 $a Vasilchuk, Daniel. $3 3191459
245 1 0 $a Novel Mechanistic Insights Into the Pressure and Temperature Stabilization of Enzymes.
300 $a 108 p.
500 $a Source: Dissertation Abstracts International, Volume: 78-02(E), Section: B.
500 $a Adviser: George I. Makhatadze.
502 $a Thesis (Ph.D.)--Rensselaer Polytechnic Institute, 2016.
520 $a Pressure and temperature have important roles in the evolution of all organisms and their associated proteins. Both factors need to be considered when attempting to engineer more stable proteins for industrial and pharmaceutical applications. In this work, the thermodynamic properties of ancestral and modern proteins were extensively characterized to determine whether the ancestral proteins had unique, or in some way improved, parameters when compared to their modern homologues. The experimental findings, with a primary focus on the pressure-related (volumetric) parameters, could provide clues for engineering more stable proteins.
520 $a Pressure perturbation calorimetry (PPC) was used to determine the volumetric properties for a diverse set of globular proteins, including six ancestral thioredoxins (Trx). The thermal expansion coefficient in the native state as a function of temperature (alphaN (T)) was shown to be very different for the studied proteins. After eliminating various structural and thermodynamic factors it was concluded that the differences in the native state expansivity function might be defined by the overall tertiary topology of the protein fold; a hypothesis that was supported by the data for the ancestral Trx. Also, the alphaN values were slightly different even for proteins with the same topology (Trx).
520 $a To investigate the thermodynamics further, a larger set of Trx was considered, comprised of ancestral and existing Trx. Differential scanning calorimetry (DSC) experiments were performed to obtain the temperature-related thermodynamic parameters and were supplemented by PPC experiments to evaluate the volumetric parameters. Pressure-temperature (P-T) profiles were constructed at equilibrium. The P-T ranges, or stability ranges, were determined by integrating the areas under these P-T profiles. The results indicate that ancestral Trx generally have improved stability ranges compared to those of modern Trx. This increase in stability range appears to originate from increased volume change upon unfolding (DeltaV). Such a novel mechanism of stabilization has not been previously derived experimentally for any other proteins. There was no correlation of stability range with the change in the thermal expansion coefficient upon unfolding (alphaDelta).
520 $a Additionally, the kinetics of folding and unfolding for the Trx were characterized to determine whether there was an optimization of kinetic properties over the evolutionary time. Based on Chevron plots, in which the log of the apparent kinetic rates is plotted against chemical denaturant concentration, we concluded that the folding kinetics were relatively conserved, but the unfolding kinetics were not. Unfolding kinetics did not correlate with evolutionary time, but correlated well with the denaturation temperature (Tm) and overall protein thermodynamic stability (DeltaG). This implied that unfolding kinetics dictate the overall stability, at least for this particular Trx fold.
520 $a Lastly, the thermodynamic parameters of a group of acylphosphatase (Acp) proteins were determined using DSC and PPC. Based on the integration of the respective P-T profiles to obtain stability ranges, a negative trend of stability range with the Deltaalpha values was determined. There was also a weaker, but positive trend of stability range with the DeltaV values. It was concluded that the Acps used a combination mechanism of stabilization by optimizing the Deltaalpha and DeltaV parameters simultaneously. A larger set of Acp variants will be needed to further confirm these biophysical trends.
590 $a School code: 0185.
650 4 $a Biochemistry. $3 518028
650 4 $a Biophysics. $3 518360
690 $a 0487
690 $a 0786
710 2 $a Rensselaer Polytechnic Institute. $b Chemistry. $3 2098707
773 0 $t Dissertation Abstracts International $g 78-02B(E).
790 $a 0185
791 $a Ph.D.
792 $a 2016
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10158621