9PVL

Crystal Structure of Fluorophore-binding Protein JF657-bp

Summary for 9PVL

• Entry DOI: 10.2210/pdb9pvl/pdb
• Descriptor: JF657-bp, 2-{2-[2-(2-{2-[2-(2-ETHOXY-ETHOXY)-ETHOXY]-ETHOXY}-ETHOXY)-ETHOXY]-ETHOXY}-ETHANOL, CALCIUM ION, ... (5 entities in total)
• Functional Keywords: de novo designed protein, small molecule binder, fluorophore-binding, de novo protein, multiplex imaging
• Biological source: synthetic construct
• Total number of polymer chains: 4
• Total formula weight: 63885.47

Authors

Bera, A.K.,Tran, L.,An, L.,Baker, D. (deposition date: 2025-08-01, release date: 2026-07-15, Last modification date: 2026-09-02)

Primary citation

Tran, L.,Klein, S.,Juergens, D.,Sharma, S.,Decarreau, J.,Lee, G.R.,Wang, Y.,Chen, W.,Bera, A.K.,Kang, A.,Woods, J.,Joyce, E.,Vafeados, D.K.,Roullier, N.,Li, X.,Liu, B.,Bo, Y.,Muratspahic, E.,Brown, T.A.,Grimm, J.B.,Patel, R.,Lavis, L.D.,Mahamid, J.,An, L.,Baker, D.
De novo design of orthogonal far-red, orange, and green fluorophore-binding proteins for multiplexed imaging.
Science, 393:813-819, 2026

PubMed Abstract: Fluorescent proteins and small-molecule dyes offer complementary advantages for biological imaging: proteins are amenable to genetic tagging, whereas dyes provide superior brightness and photostability. To combine these strengths, we used de novo protein design to generate small, nanomolar-affinity, high-selectivity binders (NovoTags) for three cell-permeable dyes spanning the visible spectrum. We show that the NovoTag fluorescent lifetimes can be tuned and demonstrate their application in lifetime and wavelength-based multiplexed fluorescence imaging. We further design a two-chain NovoTag that functions as a chemically induced dimerization system with fluorescent readout in living cells, or as a minimally perturbing proximity probe in fixed cells. Our approach combines the advantages of fluorescent proteins and small-molecule dyes, expanding the toolkit for cellular imaging.

PubMed: 42461986
DOI: 10.1126/science.aeb0822

Experimental method

X-RAY DIFFRACTION (1.44 Å)