Recovery of Red Fluorescent Protein Chromophore Maturation Deficiency through Rational Design

Moore, Matthew M.; Oteng-Pabi, Samuel K.; Pandelieva, Antonia T.; Mayo, Stephen L.; Chica, Roberto A.

doi:10.1371/journal.pone.0052463

Recovery of Red Fluorescent Protein Chromophore Maturation Deficiency through Rational Design

Journal Article · Thu Dec 20 04:00:00 EST 2012 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0052463· OSTI ID:1627573

Moore, Matthew M. ^[1]; Oteng-Pabi, Samuel K. ^[2]; Pandelieva, Antonia T. ^[2]; Mayo, Stephen L. ^[3]; Chica, Roberto A. ^[4]

California Institute of Technology (CalTech), Pasadena, CA (United States). Division of Biology; DOE/OSTI
Univ. of Ottawa, ON (Canada). Dept. of Chemistry
California Institute of Technology (CalTech), Pasadena, CA (United States). Division of Biology; California Institute of Technology (CalTech), Pasadena, CA (United States). Division of Chemistry and Chemical Engineering
Univ. of Ottawa, ON (Canada). Dept. of Chemistry; Univ. of Ottawa, ON (Canada). Centre for Catalysis Research and Innovation

Red 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.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory, Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); Gordon and Betty Moore Foundation

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1627573

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 12 Vol. 7; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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