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      Metabolic networks in motion: 13C-based flux analysis

      review-article
      1 , a
      Molecular Systems Biology
      13C experiments, fluxome, metabolic engineering, metabolic pathways, network analysis

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          Abstract

          Many properties of complex networks cannot be understood from monitoring the components—not even when comprehensively monitoring all protein or metabolite concentrations—unless such information is connected and integrated through mathematical models. The reason is that static component concentrations, albeit extremely informative, do not contain functional information per se. The functional behavior of a network emerges only through the nonlinear gene, protein, and metabolite interactions across multiple metabolic and regulatory layers. I argue here that intracellular reaction rates are the functional end points of these interactions in metabolic networks, hence are highly relevant for systems biology. Methods for experimental determination of metabolic fluxes differ fundamentally from component concentration measurements; that is, intracellular reaction rates cannot be detected directly, but must be estimated through computer model-based interpretation of stable isotope patterns in products of metabolism.

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          Most cited references92

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          Analysis of optimality in natural and perturbed metabolic networks.

          An important goal of whole-cell computational modeling is to integrate detailed biochemical information with biological intuition to produce testable predictions. Based on the premise that prokaryotes such as Escherichia coli have maximized their growth performance along evolution, flux balance analysis (FBA) predicts metabolic flux distributions at steady state by using linear programming. Corroborating earlier results, we show that recent intracellular flux data for wild-type E. coli JM101 display excellent agreement with FBA predictions. Although the assumption of optimality for a wild-type bacterium is justifiable, the same argument may not be valid for genetically engineered knockouts or other bacterial strains that were not exposed to long-term evolutionary pressure. We address this point by introducing the method of minimization of metabolic adjustment (MOMA), whereby we test the hypothesis that knockout metabolic fluxes undergo a minimal redistribution with respect to the flux configuration of the wild type. MOMA employs quadratic programming to identify a point in flux space, which is closest to the wild-type point, compatibly with the gene deletion constraint. Comparing MOMA and FBA predictions to experimental flux data for E. coli pyruvate kinase mutant PB25, we find that MOMA displays a significantly higher correlation than FBA. Our method is further supported by experimental data for E. coli knockout growth rates. It can therefore be used for predicting the behavior of perturbed metabolic networks, whose growth performance is in general suboptimal. MOMA and its possible future extensions may be useful in understanding the evolutionary optimization of metabolism.
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            Integrating high-throughput and computational data elucidates bacterial networks.

            The flood of high-throughput biological data has led to the expectation that computational (or in silico) models can be used to direct biological discovery, enabling biologists to reconcile heterogeneous data types, find inconsistencies and systematically generate hypotheses. Such a process is fundamentally iterative, where each iteration involves making model predictions, obtaining experimental data, reconciling the predicted outcomes with experimental ones, and using discrepancies to update the in silico model. Here we have reconstructed, on the basis of information derived from literature and databases, the first integrated genome-scale computational model of a transcriptional regulatory and metabolic network. The model accounts for 1,010 genes in Escherichia coli, including 104 regulatory genes whose products together with other stimuli regulate the expression of 479 of the 906 genes in the reconstructed metabolic network. This model is able not only to predict the outcomes of high-throughput growth phenotyping and gene expression experiments, but also to indicate knowledge gaps and identify previously unknown components and interactions in the regulatory and metabolic networks. We find that a systems biology approach that combines genome-scale experimentation and computation can systematically generate hypotheses on the basis of disparate data sources.
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              Just-in-time transcription program in metabolic pathways.

              A primary goal of systems biology is to understand the design principles of the transcription networks that govern the timing of gene expression. Here we measured promoter activity for approximately 100 genes in parallel from living cells at a resolution of minutes and accuracy of 10%, based on GFP and Lux reporter libraries. Focusing on the amino-acid biosynthesis systems of Escherichia coli, we identified a previously unknown temporal expression program and expression hierarchy that matches the enzyme order in unbranched pathways. We identified two design principles: the closer the enzyme is to the beginning of the pathway, the shorter the response time of the activation of its promoter and the higher its maximal promoter activity. Mathematical analysis suggests that this 'just-in-time' (ref. 5) transcription program is optimal under constraints of rapidly reaching a production goal with minimal total enzyme production. Our findings suggest that metabolic regulation networks are designed to generate precision promoter timing and activity programs that can be understood using the engineering principles of production pipelines.
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                Author and article information

                Journal
                Mol Syst Biol
                Molecular Systems Biology
                1744-4292
                2006
                14 November 2006
                : 2
                : 62
                Affiliations
                [1 ]Institute of Molecular Systems Biology, ETH Zurich, Switzerland
                Author notes
                [a ]Institute of Molecular Systems Biology, ETH Zurich, Wolfgang-Pauli-Str. 16, Zürich 8093, Switzerland. Tel.: +41 44 633 3672; Fax: +41 44 633 1051; sauer@ 123456ethz.ch
                Article
                msb4100109
                10.1038/msb4100109
                1682028
                17102807
                866fe9d9-f938-4bc1-bf8b-241a8fe0c6bc
                Copyright © 2006, EMBO and Nature Publishing Group
                History
                : 21 July 2006
                : 6 October 2006
                Page count
                Pages: 1
                Categories
                Review Article

                Quantitative & Systems biology
                fluxome,13c experiments,metabolic engineering,network analysis,metabolic pathways

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