Integrated function of a kinetic proofreading mechanism: steady-state analysis testing internal consistency of data obtained in vivo and in vitro and predicting parameter values.

Abstract:

:Experimental measurements of the kinetic mechanism involving isoleucyl-tRNA synthetase proofreading valyl-tRNAIle in Escherichia coli have been incorporated into the conventional Michaelis-Menten model for this system. The model was subjected to a detailed mathematical analysis in the steady state. The results of this analysis provide an excellent illustration of the value of integrating fragmentary data into a model of the intact system. (1) Such integration provides a rigorous test for consistency of the individual measurements. For the above synthetase system, the published experimental data were found to be internally inconsistent. (2) Such integration predicts which experimental data are most suspect. In this case, one of the three most questionable measurements, the isoleucine pool size in vivo, was found upon reexamination to be in error by 10-15-fold. Correction of this error produced a self-consistent set of parameter values. (3) The integrated analysis provides predictions for various parameter values. In many cases, these predictions provide estimates for parameter values that are difficult to determine directly or that have yet to be measured experimentally. (4) A sensitivity analysis provides an indication of the relative importance of various parameter values and, hence, an indication of where future experimental effort might be focused most profitably.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Okamoto M,Savageau MA

doi

10.1021/bi00303a019

subject

Has Abstract

pub_date

1984-04-10 00:00:00

pages

1701-9

issue

8

eissn

0006-2960

issn

1520-4995

journal_volume

23

pub_type

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