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ID 115494
Author
Mine, Masanori Tokushima University
Keywords
Capillary electrophoresis
Dynamic frontal analysis
Creatine kinase
Transphosphorylation
Kinetic analysis
Content Type
Journal Article
Description
Kinetic reactions of the transphosphorylation with creatine kinase (CK) were individually investigated between creatine (Cr) and creatine phosphate (CrP) by pressure-assisted capillary electrophoresis/dynamic frontal analysis (pCE/DFA). The transphosphorylations are reversible between Cr and CrP, and reverse reactions inevitably accompany in general batch analyses. In pCE/DFA, the kinetic reaction proceeds in a separation capillary and the product is continuously resolved from the substrate zone. Therefore, the formation rate is kept constant at the substrate zone without the reverse reaction, and the product is detected as a plateau signal. This study demonstrates the direct and individual analyses of both the forward and the backward kinetic reactions with CK by pCE/DFA. A plateau signal was detected in the pCE/DFA with ADP or ATP as one of the products on either the forward or the backward reactions. The Michaelis-Menten constants of Km,ATP (from Cr to CrP) and Km,ADP (from CrP to Cr) were successfully determined through the plateau signal. Determined values of Km,ATP and Km,ADP by pCE/DFA were smaller than the ones obtained by the pre-capillary batch analyses. The results agree with the fact that the reverse reaction is excluded in the analysis of the kinetic reactions. The proposed pCE/DFA is useful on individual analyses of both forward and backward kinetic reactions without any interference from the reverse reaction.
Journal Title
Analytical and Bioanalytical Chemistry
ISSN
16182642
16182650
NCID
AA11610734
Publisher
Springer Nature
Volume
413
Issue
5
Start Page
1453
End Page
1460
Published Date
2021-01-21
Remark
This is a post-peer-review, pre-copyedit version of an article published in Analytical and Bioanalytical Chemistry. The final authenticated version is available online at: http://dx.doi.org/10.1007/s00216-020-03110-9.
論文本文は2022-01-21以降公開予定
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
language
eng
TextVersion
その他
departments
Science and Technology