東華大學圖書館 |

Language: English

Help

回圖書館首頁

手機版館藏查詢

Back

Switch To: Labeled | MARC Mode | ISBD

Measurement of Kinetic Isotope Effec...

Fernandez, Patrick Lista,

Linked to FindBook

Google Book

Amazon

博客來

Measurement of Kinetic Isotope Effects by Dimer Distribution Analysis at Reaction Completion: Theory and Application to D-Alanine:D-Alanine Ligase /

Record Type:	Electronic resources : Monograph/item
Title/Author:	Measurement of Kinetic Isotope Effects by Dimer Distribution Analysis at Reaction Completion: Theory and Application to D-Alanine:D-Alanine Ligase // Patrick Lista Fernandez.
Author:	Fernandez, Patrick Lista,
Description:	1 electronic resource (327 pages)
Notes:	Source: Dissertations Abstracts International, Volume: 82-04, Section: B.
Contained By:	Dissertations Abstracts International82-04B.
Subject:	Biochemistry. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=28090982
ISBN:	9798672198354

Measurement of Kinetic Isotope Effects by Dimer Distribution Analysis at Reaction Completion: Theory and Application to D-Alanine:D-Alanine Ligase /
Fernandez, Patrick Lista,

Measurement of Kinetic Isotope Effects by Dimer Distribution Analysis at Reaction Completion: Theory and Application to D-Alanine:D-Alanine Ligase /Patrick Lista Fernandez. - 1 electronic resource (327 pages)

Source: Dissertations Abstracts International, Volume: 82-04, Section: B.

The stoichiometric requirement of two of the same substrate per reaction in the dimerization catalyzed by Mycobacterium tuberculosis D-alanine:D-alanine ligase (Ddl) is a feature less commonly encountered in enzyme-catalyzed reactions. The cumulative account of isotopic discrimination for the monomers at the transition state is encrypted by their subsequent placement in the dipeptide termini at reaction completion. The theoretical framework for mining KIE information through this unique phenomenon was formalized and was applied to the dipeptide mixture resulting from the internal competition of unlabeled and 13C-labeled D-alanines by combining NMR spectroscopy and mass spectrometry. The 13C-KIEs measured for the acyl- (1.011 ± 0.004) and alpha- (1.009 ± 0.011) positions, corroborated by the alpha 2H-KIEs, i.e., D(kcat/Km) = 1.0 ± 0.3 and Dkcat = 0.99 ± 0.08, are consistent with the partially rate-limiting collapse of the tetrahedral intermediate with an early transition state.In contrast to dimerization, dipeptide hydrolysis was found to be highly unfavorable due to substrate (appKi, pH 7.8 = 4.9 ± 0.9 mM) and product (appKi, pH 8 ≈ 0.1 mM) inhibition. However, with a balanced symphony of coupling enzymes, ad libitum O2 access was able to orchestrate the reaction to proceed irreversibly. These results, in addition to the synthesis of 13C-labeled D-alanine-D-alanine through stereoselective and robust chemistries, establish the groundwork for the measurement of KIEs in the cleavage direction.

English

ISBN: 9798672198354Subjects--Topical Terms:

518028
Biochemistry.
Subjects--Index Terms:

Dimerization

Measurement of Kinetic Isotope Effects by Dimer Distribution Analysis at Reaction Completion: Theory and Application to D-Alanine:D-Alanine Ligase /
LDR:03124nmm a22004573i 4500 001 2437592
005 20251124054943.5
006 m o d
007 cr|nu||||||||
008 261011s2020 miu||||||m |||||||eng d
020 $a 9798672198354
035 $a (MiAaPQD)AAI28090982
035 $a AAI28090982
040 $a MiAaPQD $b eng $c MiAaPQD $e rda
100 1 $a Fernandez, Patrick Lista, $e author. $0 (orcid)0000-0002-1862-5063 $3 3828638
245 1 0 $a Measurement of Kinetic Isotope Effects by Dimer Distribution Analysis at Reaction Completion: Theory and Application to D-Alanine:D-Alanine Ligase / $c Patrick Lista Fernandez.
264 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2020
300 $a 1 electronic resource (327 pages)
336 $a text $b txt $2 rdacontent
337 $a computer $b c $2 rdamedia
338 $a online resource $b cr $2 rdacarrier
500 $a Source: Dissertations Abstracts International, Volume: 82-04, Section: B.
500 $a Advisors: Murkin, Andrew Committee members: Morrow, Janet; Diver, Steven; Cook, Timothy.
502 $b Ph.D. $c State University of New York at Buffalo $d 2020.
520 $a The stoichiometric requirement of two of the same substrate per reaction in the dimerization catalyzed by Mycobacterium tuberculosis D-alanine:D-alanine ligase (Ddl) is a feature less commonly encountered in enzyme-catalyzed reactions. The cumulative account of isotopic discrimination for the monomers at the transition state is encrypted by their subsequent placement in the dipeptide termini at reaction completion. The theoretical framework for mining KIE information through this unique phenomenon was formalized and was applied to the dipeptide mixture resulting from the internal competition of unlabeled and 13C-labeled D-alanines by combining NMR spectroscopy and mass spectrometry. The 13C-KIEs measured for the acyl- (1.011 ± 0.004) and alpha- (1.009 ± 0.011) positions, corroborated by the alpha 2H-KIEs, i.e., D(kcat/Km) = 1.0 ± 0.3 and Dkcat = 0.99 ± 0.08, are consistent with the partially rate-limiting collapse of the tetrahedral intermediate with an early transition state.In contrast to dimerization, dipeptide hydrolysis was found to be highly unfavorable due to substrate (appKi, pH 7.8 = 4.9 ± 0.9 mM) and product (appKi, pH 8 ≈ 0.1 mM) inhibition. However, with a balanced symphony of coupling enzymes, ad libitum O2 access was able to orchestrate the reaction to proceed irreversibly. These results, in addition to the synthesis of 13C-labeled D-alanine-D-alanine through stereoselective and robust chemistries, establish the groundwork for the measurement of KIEs in the cleavage direction.
546 $a English
590 $a School code: 0656
650 4 $a Biochemistry. $3 518028
650 4 $a Pharmaceutical sciences. $3 3173021
650 4 $a Chemistry. $3 516420
653 $a Dimerization
653 $a Enzymology
653 $a Kinetic isotope effects
653 $a Kinetics
653 $a Transition state
653 $a Mycobacterium tuberculosis
653 $a D-alanine:D-alanine
690 $a 0487
690 $a 0572
690 $a 0485
710 2 $a State University of New York at Buffalo. $b Chemistry. $e degree granting institution. $3 3824228
720 1 $a Murkin, Andrew $e degree supervisor.
773 0 $t Dissertations Abstracts International $g 82-04B.
790 $a 0656
791 $a Ph.D.
792 $a 2020
856 4 0 $u https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=28090982