6UBO

Fluorogen Activating Protein Dib1

Summary for 6UBO

• Entry DOI: 10.2210/pdb6ubo/pdb
• Descriptor: Outer membrane lipoprotein Blc, 12-(diethylamino)-2,2-bis(fluoranyl)-4,5-dimethyl-5-aza-3-azonia-2-boranuidatricyclo[7.4.0.0^{3,7}]trideca-1(13),3,7,9,11-pentaen-6-one, CITRIC ACID, ... (5 entities in total)
• Functional Keywords: fluorogen activating protein, lipocalin, biomarker, membrane protein, transport protein
• Biological source: Escherichia coli (strain K12)
• Total number of polymer chains: 2
• Total formula weight: 42743.39

Authors

Muslinkina, L.,Pletneva, N.,Pletnev, V.Z.,Pletnev, S. (deposition date: 2019-09-12, release date: 2020-09-16, Last modification date: 2023-10-11)

Primary citation

Muslinkina, L.,Gavrikov, A.S.,Bozhanova, N.G.,Mishin, A.S.,Baranov, M.S.,Meiler, J.,Pletneva, N.V.,Pletnev, V.Z.,Pletnev, S.
Structure-Based Rational Design of Two Enhanced Bacterial Lipocalin Blc Tags for Protein-PAINT Super-resolution Microscopy.
Acs Chem.Biol., 15:2456-2465, 2020

PubMed Abstract: Super-resolution fluorescent imaging in living cells remains technically challenging, largely due to the photodecomposition of fluorescent tags. The recently suggested protein-PAINT is the only super-resolution technique available for prolonged imaging of proteins in living cells. It is realized with complexes of fluorogen-activating proteins, expressed as fusions, and solvatochromic synthetic dyes. Once photobleached, the dye in the complex is replaced with a fresh fluorogen available in the sample. With suitable kinetics, this replacement creates fluorescence blinking required for attaining super-resolution and overcomes photobleaching associated with the loss of an irreplaceable fluorophore. Here we report on the rational design of two protein-PAINT tags based on the 1.58 Å crystal structure of the DiB1:M739 complex, an improved green-emitting DiB3/F74V:M739 and a new orange-emitting DiB3/F53L:M739. They outperform previously reported DiB-based tags to become best in class biomarkers for protein-PAINT. The new tags advance protein-PAINT from the proof-of-concept to a reliable tool suitable for prolonged super-resolution imaging of intracellular proteins in fixed and living cells and two-color PAINT-like nanoscopy with a single fluorogen.

PubMed: 32809793
DOI: 10.1021/acschembio.0c00440

Experimental method

X-RAY DIFFRACTION (1.58 Å)