Deciphering metabolic networks by blue native polyacrylamide gel electrophoresis: A functional proteomic exploration

• BN-PAGE is applied to separate proteins in native form.
• Enzymatic activity assays in metabolic networks can be performed following electrophoresis.
• BN-PAGE maintains in vivo protein–protein interactions.
• Novel protein interactions can be elucidated via secondary analyses.
• Inter and intra-organellar metabolic networks can be uncovered by BN-PAGE.

Metabolism is the consortium of reactions within a cell which directs a variety of processes including energy synthesis, signalling and the behaviour of a biological system. Metabolic networks, and more specifically the activity of enzymes within them, provide an accurate status of how cellular information is being executed. The performance of these networks and their ability to siphon metabolites in a number of directions may be the difference between a healthy and diseased state. Blue native polyacrylamide gel electrophoresis (BN-PAGE), owing to its simplicity and wide-ranging applications, permits the inspection of these nodules. The separation of proteins and enzyme complexes in their native format enables the exploration of enzymatic activity in metabolic networks via in-gel assays. These are quick, specific, and amenable to further studies. This electrophoretic technology not only enables the visualization of enzymatic efficacy but reveals the crosstalk among enzymes and their interactions with other organellar partners.

Figure optionsDownload as PowerPoint slide