A normalized plot as a novel and time-saving tool in complex enzyme kinetic analysis

被引:8
作者
Bravo, IG [1 ]
Busto, F [1 ]
De Arriaga, D [1 ]
Ferrero, MA [1 ]
Rodríguez-Aparicio, LB [1 ]
Martínez-Blanco, H [1 ]
Reglero, A [1 ]
机构
[1] Univ Leon, Dept Bioquim & Biol Mol, E-24071 Leon, Spain
关键词
data normalization; enzymology; model-fitting; parameter estimation; rate equations;
D O I
10.1042/bj3580573
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A new data treatment is described for designing kinetic experiments and analysing kinetic results for multi-substrate enzymes. Normalized velocities are plotted against normalized substrate concentrations. Data are grouped into n+1 families across the range of substrate or product tested, n being the number of substrates plus products assayed. It has the following advantages over traditional methods: (1) it reduces to less than a half the amount of data necessary for a proper description of the system; (2) it introduces a self-consistency checking parameter that ensures the 'scientific reliability' of the mathematical output; (3) it eliminates the need for a prior knowledge of V-max; (4) the normalization of data allows the use of robust and fuzzy methods suitable for managing really 'noisy' data; (5) it is appropriate for analysing complex systems, as the complete general equation is used, and the actual influence of effectors can be typified; (6) it is amenable to being implemented as a software that incorporates testing and electing among rival kinetic models.
引用
收藏
页码:573 / 583
页数:11
相关论文
共 18 条
[1]  
BEECHEM JM, 1992, METHOD ENZYMOL, V210, P37
[2]   Kinetic properties of the acylneuraminate cytidylyltransferase from Pasteurella haemolytica A2 [J].
Bravo, IG ;
Barrallo, S ;
Ferrero, MA ;
Rodriguez-Aparicio, LB ;
Martínez-Blanco, H ;
Reglero, A .
BIOCHEMICAL JOURNAL, 2001, 358 (358) :585-598
[3]   COMBINATION PLOTS AS GRAPHICAL TOOLS IN THE STUDY OF ENZYME-INHIBITION [J].
CHAN, WWC .
BIOCHEMICAL JOURNAL, 1995, 311 :981-985
[4]  
Cleland W W, 1979, Methods Enzymol, V63, P103
[5]  
CORNISHBOWDEN A, 1995, FUNDAMENTALS ENZYME, P93
[6]  
CORNISHBOWDEN A, 1995, ANAL ENZYME KINETIC, P3
[7]  
DICERA E, 1992, METHOD ENZYMOL, V210, P68
[8]  
Fromm HJ, 1975, INITIAL RATE ENZYME
[9]  
HUNTER A, 1945, J BIOL CHEM, V157, P427
[10]  
Johnson ML, 2000, METHOD ENZYMOL, V321, P424