Metabolic control, experimental analysis

Fell, D.A. (1992). Metabolic control analysis A survey of its theoretical and experimental development. Biochem. J. 286,313-330. 

Metabolic control analysis (MCA) assigns a flux control coefficient (FCC) to each step in the pathway and considers the sum of the coefficients. Competing pathway components may have negative FCCs. To measure FCCs, a variety of experimental techniques including radio isotopomers and pulse chase experiments are necessary in a tissue culture system. Perturbation of the system, for example, with over-expression of various genes can be applied iteratively to understand and optimize product accumulation. 

Martens, D. E. Hugenholtz, J. Kleere-bezem, M. et al. Metabolic engineering of lactic add bacteria, the combined approach kinetic modelling, metabolic control and experimental analysis. Microbiology 2002,148 1003-1013. 

Distributive Control of Metabolism Experimental Analysis of Metabolism Goals for the Study of Metabolism... 

The wide gap between the two opposing theories, replication first and metabolism first , was analysed by Pross from the Ben Gurion University of the Negev (Israel). Pross concludes that replication came first He is convinced that a causality between the two theories can only be established if it is assumed that the replication-first thesis is correct. His analysis also shows that more of the experimental results and theoretical rationales favour the replication thesis. The author finds his assumption justified that life processes are strongly kinetically controlled and that the development of metabolic pathways can only be understood if life is considered as a manifestation of replicative chemistry (Pross, 2004). 

Based on the accessibility of high-quality experimental information, we now focus on aspects of model interrogation and analysis. The question how cells actually control and distribute their flux under different conditions requires a mathematical and formal approach to metabolic regulation. The knowledge obtained by quantitative experiments must be, in the sense of Section II.B, encoded into a mathematical system, scrutinized utilizing the tools of formal analysis, and eventually decoded back into predictions about the natural system. 

Figure 4.10. Proteomic analysis by SILAC. Proteomic analysis by SILAC or stable isotope labeling of amino acids in cell culture utilize de novo metabolic incorporation of stable-isotope-labeled amino acids during protein synthesis. Cells can be cultured with various combinations of stable-isotope-labeled amino acids such as lysine or arginine. Tyrosine has been used in phosphoprotein studies of tyrosine residues. About five or six cell divisions are needed for complete labeling of proteins in cell cultures prior to experimentation. Labeled cells from control and treatment(s) lysates are combined and digested. Quantitation and identification are performed by LC-MS/MS.

---