substrate cooperativity in marine luciferases在海洋荧光素酶底物协同.pdfVIP

Substrate Cooperativity in Marine Luciferases 1 2 2 George Tzertzinis , Ezra Schildkraut , Ira Schildkraut * 1 New England Biolabs, Inc., Ipswich, Massachusetts, United States of America, 2 CerroSci LLC, Ipswich, Massachusetts, United States of America Abstract Marine luciferases are increasingly used as reporters to study gene regulation. These luciferases have utility in bioluminescent assay development, although little has been reported on their catalytic properties in response to substrate concentration. Here, we report that the two marine luciferases from the copepods, Gaussia princeps (GLuc) and Metridia longa (MLuc) were found, surprisingly, to produce light in a cooperative manner with respect to their luciferin substrate concentration; as the substrate concentration was decreased 10 fold the rate of light production decreased 1000 fold. This positive cooperative effect is likely a result of allostery between the two proposed catalytic domains found in Gaussia and Metridia. In contrast, the marine luciferases from Renilla reniformis (RLuc) and Cypridina noctiluca (CLuc) demonstrate a linear relationship between the concentration of their respective luciferin and the rate of light produced. The consequences of these enzyme responses are discussed. Citation: Tzertzinis G, Schildkraut E, Schildkraut I (2012) Substrate Cooperativity in Marine Luciferases. PLoS ONE 7(6): e40099. doi:10.1371/journal.pone.0040099 Editor: Subba Reddy Palli, U. Kentucky, United States of America Received May 10, 2012; Accepted May 31, 2012; Published June 29, 2012 Copyright: 2012 Tzertzinis et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribut