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Stoichiometry Of The Α‑complementation Reaction Of Escherichia Coli Β‑galactosidase As Revealed Through Single-molecule Studies

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Biochemistry | February 10, 2015

Pratyusha Mogalisetti and David R. Walt*
DOI: 10.1021/bi5015024

Abstract: The α-complementation reaction of β-galactosidase was studied at single-molecule resolution using arrays of femtoliter-sized wells. Single molecules of the complementation species were observed to be stable for long periods of time, demonstrating that the α-complementation reaction is irreversible. By directly counting the number of active molecules formed in the complementation reaction when different concentrations of enzyme acceptor (EA) and enzyme donor (ED) are used, we deduce that the EA:ED ratio in the complementation species is 4:1.