Recovery of Red Fluorescent Protein Chromophore Maturation Deficiency through Rational Design

FieldValue
dc.contributor.authorMoore, Matthew M.
dc.contributor.authorPandelieva, Antonia T.
dc.contributor.authorMayo, Stephen L.
dc.contributor.authorChica, Roberto A.
dc.contributor.authorOteng-Pabi, Samuel K.
dc.date.accessioned2013-03-05T20:30:06Z
dc.date.available2013-03-05T20:30:06Z
dc.date.created2012
dc.date.issued2012
dc.identifier.urihttp://hdl.handle.net/10393/23910
dc.identifier.urihttp://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0052463
dc.description.abstractRed fluorescent proteins (RFPs) derived from organisms in the class Anthozoa have found widespread application as imaging tools in biological research. For most imaging experiments, RFPs that mature quickly to the red chromophore and produce little or no green chromophore are most useful. In this study, we used rational design to convert a yellow fluorescent mPlum mutant to a red-emitting RFP without reverting any of the mutations causing the maturation deficiency and without altering the red chromophore’s covalent structure. We also created an optimized mPlum mutant (mPlum-E16P) that matures almost exclusively to the red chromophore. Analysis of the structure/function relationships in these proteins revealed two structural characteristics that are important for efficient red chromophore maturation in DsRed-derived RFPs. The first is the presence of a lysine residue at position 70 that is able to interact directly with the chromophore. The second is an absence of non-bonding interactions limiting the conformational flexibility at the peptide backbone that is oxidized during red chromophore formation. Satisfying or improving these structural features in other maturation-deficient RFPs may result in RFPs with faster and more complete maturation to the red chromophore.
dc.language.isoen
dc.titleRecovery of Red Fluorescent Protein Chromophore Maturation Deficiency through Rational Design
dc.typeArticle
dc.identifier.doi10.1371/journal.pone.0052463
CollectionChimie et sciences biomoléculaires // Chemistry and Biomolecular Sciences
Publications en libre accès financées par uOttawa // uOttawa financed open access publications

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