Improvements in metabolic flux analysis using carbon bond labeling experiments: bondomer balancing and Boolean function mapping.
The biosynthetically directed fractional (13)C labeling method for metabolic flux evaluation relies on performing a 2-D [(13)C, (1)H] NMR experiment on extracts from organisms cultured on a uniformly labeled carbon substrate. This article focuses on improvements in the interpretation of data obtained from such an experiment by employing the concept of bondomers. Bondomers take into account the natural abundance of (13)C; therefore many bondomers in a real network are zero, and can be precluded a priori--thus resulting in fewer balances. Using this method, we obtained a set of linear equations which can be solved to obtain analytical formulas for NMR-measurable quantities in terms of fluxes in glycolysis and the pentose phosphate pathways. For a specific case of this network with four degrees of freedom, a priori identifiability of the fluxes was shown possible for any set of fluxes. For a more general case with five degrees of freedom, the fluxes were shown identifiable for a representative set of fluxes. Minimal sets of measurements which best identify the fluxes are listed. Furthermore, we have delineated Boolean function mapping, a new method to iteratively simulate bondomer abundances or efficiently convert carbon skeleton rearrangement information to mapping matrices. The efficiency of this method is expected to be valuable while analyzing metabolic networks which are not completely known (such as in plant metabolism) or while implementing iterative bondomer balancing methods.
Sriram
Ganesh
G
Department of Chemical Engineering, Iowa State University, 3031, Sweeney Hall, Ames, IA 50011-2230, USA.
Shanks
Jacqueline V
JV
D000465:Algorithms; D001665:Binding Sites; D002247:Carbon Isotopes / Q000378:metabolism; D003198:Computer Simulation; D006019:Glycolysis / Q000502:physiology*; D007553:Isotope Labeling / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008660:Metabolism / Q000502:physiology*; D008954:Models, Biological; D010427:Pentose Phosphate Pathway / Q000502:physiology*
D002247:Carbon Isotopes