3M24

Crystal structure of TagBFP fluorescent protein

Summary for 3M24

• Entry DOI: 10.2210/pdb3m24/pdb
• Related: 3M22
• Descriptor: TagBFP, 2,3-DIHYDROXY-1,4-DITHIOBUTANE, GLYCEROL, ... (5 entities in total)
• Functional Keywords: acylimine-containing blue and red chromophores, fluorescent proteins, de novo protein
• Biological source: synthetic construct (artificial gene)
• Total number of polymer chains: 4
• Total formula weight: 107028.49

Authors

Malashkevich, V.N.,Subach, O.M.,Almo, S.C.,Verkhusha, V.V. (deposition date: 2010-03-06, release date: 2010-05-26, Last modification date: 2023-11-15)

Primary citation

Subach, O.M.,Malashkevich, V.N.,Zencheck, W.D.,Morozova, K.S.,Piatkevich, K.D.,Almo, S.C.,Verkhusha, V.V.
Structural characterization of acylimine-containing blue and red chromophores in mTagBFP and TagRFP fluorescent proteins.
Chem.Biol., 17:333-341, 2010

PubMed Abstract: We determined the 2.2 A crystal structures of the red fluorescent protein TagRFP and its derivative, the blue fluorescent protein mTagBFP. The crystallographic analysis is consistent with a model in which TagRFP has the trans coplanar anionic chromophore with the conjugated pi-electron system, similar to that of DsRed-like chromophores. Refined conformation of mTagBFP suggests the presence of an N-acylimine functionality in its chromophore and single C(alpha)-C(beta) bond in the Tyr64 side chain. Mass spectrum of mTagBFP chromophore-bearing peptide indicates a loss of 20 Da upon maturation, whereas tandem mass spectrometry reveals that the C(alpha)-N bond in Leu63 is oxidized. These data indicate that mTagBFP has a new type of the chromophore, N-[(5-hydroxy-1H-imidazole-2-yl)methylidene]acetamide. We propose a chemical mechanism in which the DsRed-like chromophore is formed via the mTagBFP-like blue intermediate.

PubMed: 20416505
DOI: 10.1016/j.chembiol.2010.03.005

Experimental method

X-RAY DIFFRACTION (2.2 Å)