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Papers In Press, published online ahead of print December 31, 2001
J. Biol. Chem, 10.1074/jbc.C100694200
Submitted on December 3, 2001
Revised on December 28, 2001
Accepted on December 31, 2001
Pathology, Stanford University, Stanford, CA 94305
Corresponding Author: terskikh{at}stanford.edu
Earlier mutagenesis of the red fluorescent protein drFP583, also called DsRed, resulted in a mutant named Fluorescent Timer1. Further mutagenesis generated variants with novel and improved fluorescent properties. The mutant called AG4, exhibits only green fluorescence. The mutant called E5up (V105A), shows complete fluorophore maturation, eventually eliminating residual green fluorescence, present in DsRed. Finally, the mutant called E57 (V105A, I161T, S197A), matures faster than DsRed as demonstrated in vitro with purified protein and in vivo with recombinant protein expressed in E.coli and Xenopus leavis. Comparative analysis of the mutants in the context of the crystal structure of DsRed suggests that mutants with free space around the fluorophore mature faster and more completely.
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