Structural basis for the fast maturation of Arthropoda green fluorescent protein.
Since the cloning of Aequorea victoria green fluorescent protein (GFP) in 1992, a family of known GFP-like proteins has been growing rapidly. Today, it includes more than a hundred proteins with different spectral characteristics cloned from Cnidaria species. For some of these proteins, crystal structures have been solved, showing diversity in chromophore modifications and conformational states. However, we are still far from a complete understanding of the origin, functions and evolution of the GFP family. Novel proteins of the family were recently cloned from evolutionarily distant marine Copepoda species, phylum Arthropoda, demonstrating an extremely rapid generation of fluorescent signal. Here, we have generated a non-aggregating mutant of Copepoda fluorescent protein and solved its high-resolution crystal structure. It was found that the protein beta-barrel contains a pore, leading to the chromophore. Using site-directed mutagenesis, we showed that this feature is critical for the fast maturation of the chromophore.
Evdokimov
Artem G
AG
X-ray crystallography, HCRC, Discovery, Procter&Gamble Pharmaceuticals, 8700 Mason-Montgomery Road, Mason, Ohio 45040, USA.
Pokross
Matthew E
ME
Egorov
Nikolay S
NS
Zaraisky
Andrey G
AG
Yampolsky
Ilya V
IV
Merzlyak
Ekaterina M
EM
Shkoporov
Andrey N
AN
Sander
Ian
I
Lukyanov
Konstantin A
KA
Chudakov
Dmitriy M
DM
D000818:Animals; D001181:Arthropods / Q000737:chemistry* / Q000378:metabolism; D033342:Copepoda / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray / Q000379:methods; D004625:Embryo, Nonmammalian; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D014176:Protein Biosynthesis; D011487:Protein Conformation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D040681:Structural Homology, Protein; D014982:Xenopus laevis / Q000196:embryology
D011993:Recombinant Fusion Proteins; C485192:green fluorescent protein, Aequorea victoria; D049452:Green Fluorescent Proteins