%0 Journal Article
%A Zagranichny, Vasily E.
%A Rudenko, Natalia V.
%A Gorokhovatsky, Andrey Yu.
%A Zakharov, Mikhail V.
%A Balashova, Tamara A.
%A Arseniev, Alexander S.
%D 2004
%T Traditional GFP-Type Cyclization and Unexpected Fragmentation Site in a Purple
Chromoprotein from Anemonia sulcata, asFP595†
%U https://acs.figshare.com/articles/journal_contribution/Traditional_GFP_Type_Cyclization_and_Unexpected_Fragmentation_Site_in_a_Purple_Chromoprotein_from_i_Anemonia_sulcata_i_asFP595_sup_sup_/3319261
%R 10.1021/bi0488247.s001
%2 https://ndownloader.figshare.com/files/5158027
%K polypeptide chain cleavage
%K asFP 595 chromopeptides
%K Anemonia sulcata
%K unsubstituted imino moiety
%K asFP 595 chromophore
%K Met
%K proteolysis products
%K asFP 595
%K NMR
%K asFP 595 chromophore structure
%K Unexpected Fragmentation Site
%K DsRed subfamily
%K protein
%K GFP
%X The purple chromoprotein (asFP595) from Anemonia sulcata belongs to the family of green
fluorescent protein (GFP). Absorption and emission spectra of asFP595 are similar to those of a number
of recently cloned GFP-like red proteins of the DsRed subfamily. The earlier proposed asFP595
chromophore structure [Martynov, V. I.; et al. (2001) J. Biol. Chem. 276, 21012−21016] was postulated
to result from an “alternative cyclization” giving rise to a pyrazine-type six-membered heterocycle. Here
we report that the asFP595 chromophore is actually very close in chemical structure to that of zFP538,
a yellow fluorescent protein [Zagranichny, V. E.; et al. (2004) Biochemistry 43, 4764−4772]. NMR
spectroscopic studies of four chromophore-containing peptides (chromopeptides) isolated under mild
conditions from enzymatic digests of asFP595 and one chromopeptide obtained from DsRed revealed
that all of them contain a p-hydroxybenzylideneimidazolinone moiety formed by Met-65/Gln-66, Tyr-66/67, and Gly-67/68 of asFP595/DsRed, respectively. Two asFP595 chromopeptides are proteolysis
products of an isolated full-length polypeptide containing a GFP-type chromophore already formed and
arrested at an earlier stage of maturation. The two other asFP595 chromopeptides were isolated as proteolysis
products of the purified chromophore-containing C-terminal fragment. One of these has an oxo group at
Met-65 Cα and is a hydrolysis product of another one, with the imino group at Met-65 Cα. The
N-unsubstituted imino moiety of the latter is generated by spontaneous polypeptide chain cleavage at a
very unexpected site, the former peptide bond between Cys-64 C‘ and Met-65 Nα. Our data strongly
suggest that both zFP538 and asFP595 could be attributed to the DsRed subfamily of GFP-like proteins.
%I ACS Publications