Monitoring substrate binding

In principle, the double-mixing approach can be used to monitor the binding of any substrate to extracted... 

Important results are obtained in an experiment where enzyme is mixed with an excess of substrate and then the formation of product is monitored over the time period of a few enzyme turnovers and with an enzyme concentration high enough to allow quantification of one product per enzyme site. Under conditions in which substrate binding and chemical conversion to product are faster than the release of product, one observes a burst of product formation equal to one product per enzyme site, followed by steady-state turnover. 

Monitoring the substrate binding to the MBP-GalT array on SPR sensor chip... 

Fukamizo, T., Dennhart, N., Taira, T., Skriver, K., and Letzel, T. 2008. Mass spectrometric analysis of the reaction catalyzed by chitinases Real-time monitoring of the reaction, substrate binding in gas phase, and inhibition studies. Abstract paper of the 24th International Carbohydrate Symposium, Oslo, Norway, G-O002. 

To monitor the substrate binding and species present during catalysis, mass spectrometry and Cd NMR spectroscopy were conducted with mixtures of the complex with phosphoester substrates (Fig. 5.10). 

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