Amperometric monitoring of redox activity in living yeast cells:: comparison of menadione and menadione sodium bisulfite as electron transfer mediators

被引:57
作者
Heiskanen, A
Yakovleva, J
Spégel, C
Taboryski, R
Koudelka-Hep, M
Emnéus, J
Ruzgas, T
机构
[1] Lund Univ, Dept Analyt Chem, SE-22100 Lund, Sweden
[2] Soph Biosci AS, DK-2750 Ballerup, Denmark
[3] Univ Neuchatel, Inst Microtechnol, CH-2007 Neuchatel, Switzerland
关键词
amperometric method; living cells; NAD(P)H oxidizing enzymes; artificial electron acceptors; real-time;
D O I
10.1016/j.elecom.2003.12.003
中图分类号
O646 [电化学、电解、磁化学];
学科分类号
081704 ;
摘要
An amperometric method was applied for real-time monitoring of intracellular redox enzyme activity. Baker's yeast (Saccharomyces cerevisiae) cells were immobilized on platinum microband electrodes and mediated anodic currents were measured. The currents were observed in the absence and in the presence of glucose as a source of reducing equivalents, NADH and NADPH. 2-Methyl-1,4-naphthoquinone (menadione, vitamin K-3) and water soluble 2-methyl-1,4-naphthoquinone sodium bisulfite (menadione sodium bisulfite MSB) were compared as artificial electron acceptors for their ability to transduce internal cellular redox activity into electrode current. It was found that hydrophobic menadione was superior to its water-soluble bisulfite derivative for probing whole intact cells. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:219 / 224
页数:6
相关论文
共 20 条
[1]   Analysis of individual biochemical events based on artificial synapses using ultramicroelectrodes: cellular oxidative burst [J].
Amatore, C ;
Arbault, S ;
Bruce, D ;
de Oliveira, P ;
Erard, M ;
Vuillaume, M .
FARADAY DISCUSSIONS, 2000, 116 :319-333
[2]   Detection of two distinct substrate-dependent catabolic responses in yeast cells using a mediated electrochemical method [J].
Baronian, KHR ;
Downard, AJ ;
Lowen, RK ;
Pasco, N .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2002, 60 (1-2) :108-113
[3]   Scanning electrochemical microscopy of living cells.: 3.: Rhodobacter sphaeroides [J].
Cai, CX ;
Liu, B ;
Mirkin, MV ;
Frank, HA ;
Rusling, JF .
ANALYTICAL CHEMISTRY, 2002, 74 (01) :114-119
[4]   Preparation of alginate/galactosylated chitosan scaffold for hepatocyte attachment [J].
Chung, TW ;
Yang, J ;
Akaike, T ;
Cho, KY ;
Nah, JW ;
Kim, SI ;
Cho, CS .
BIOMATERIALS, 2002, 23 (14) :2827-2834
[5]   Persuasive evidence that quinone reductase type 1 (DT diaphorase) protects cells against the toxicity of electrophiles and reactive forms of oxygen [J].
Dinkova-Kostova, LT ;
Talalay, P .
FREE RADICAL BIOLOGY AND MEDICINE, 2000, 29 (3-4) :231-240
[6]   Ferricyanide reduction by Escherichia coli:: Kinetics, mechanism, and application to the optimization of recombinant fermentations [J].
Ertl, P ;
Unterladstaetter, U ;
Bayer, K ;
Mikkelsen, SR .
ANALYTICAL CHEMISTRY, 2000, 72 (20) :4949-4956
[7]  
Fiaccabrino GC, 1998, ELECTROANAL, V10, P217, DOI 10.1002/(SICI)1521-4109(199804)10:4<217::AID-ELAN217>3.0.CO
[8]  
2-W
[9]   Measurement of dicumarol-sensitive NADPH:(menadione cytochrome c) oxidoreductase activity results in an artifactual assay of DT-diaphorase in cell sonicates [J].
Hodnick, WF ;
Sartorelli, AC .
ANALYTICAL BIOCHEMISTRY, 1997, 252 (01) :165-168
[10]   Yeast and drug discovery [J].
Hughes T.R. .
Functional & Integrative Genomics, 2002, 2 (4-5) :199-211