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. 2001 Sep 15;358(Pt 3):573–583. doi: 10.1042/bj3580573

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

I G Bravo 1, F Busto 1, D De Arriaga 1, M A Ferrero 1, L B Rodríguez-Aparicio 1, H Martínez-Blanco 1, A Reglero 1
PMCID: PMC1222113  PMID: 11577687

Abstract

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 Vmax; (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.

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Selected References

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