Structural rearrangements near the chromophore influence the maturation speed and brightness of DsRed variants

  • Strongin D
  • Bevis B
  • Khuong N
 et al. 
  • 81

    Readers

    Mendeley users who have this article in their library.
  • 37

    Citations

    Citations of this article.

Abstract

The red fluorescent protein DsRed has been extensively engineered for use as an in vivo research tool. In fast maturing DsRed variants, the chromophore maturation half-time is approximately 40 min, compared to approximately 12 h for wild-type DsRed. Further, DsRed has been converted from a tetramer into a monomer, a task that entailed mutating approximately 20% of the amino acids. These engineered variants of DsRed have proven extremely valuable for biomedical research, but the structural basis for the improved characteristics has not been thoroughly investigated. Here we present a 1.7 A crystal structure of the fast maturing tetrameric variant DsRed.T4. We also present a biochemical characterization and 1.6 A crystal structure of the monomeric variant DsRed.M1, also known as DsRed-Monomer. Analysis of the crystal structures suggests that rearrangements of Ser69 and Glu215 contribute to fast maturation, and that positioning of the Lys70 side chain modulates fluorescence quantum yield. Despite the 45 mutations in DsRed.M1 relative to wild-type DsRed, there is a root-mean-square deviation of only 0.3 A between the two structures. We propose that novel intramolecular interactions in DsRed.M1 partially compensate for the loss of intermolecular interactions found in the tetramer.

Author-supplied keywords

  • DsRed
  • Fast maturation
  • Fluorescent protein
  • Monomeric
  • Structure

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Get full text

Authors

  • Daniel E. Strongin

  • Brooke Bevis

  • Nhi Khuong

  • Maureen E. Downing

  • Rita L. Strack

  • Karthik Sundaram

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free