Traditional GFP-Type Cyclization and Unexpected Fragmentation Site in a Purple Chromoprotein from Anemonia sulcata, asFP595
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- 作者：Vasily E. Zagranichny ; Natalia V. Rudenko ; Andrey Yu. Gorokhovatsky ; Mikhail V. Zakharov ; Tamara A. Balashova ; and Alexander S. Arseniev
- 刊名：Biochemistry
- 出版年：2004
- 出版时间：October 26, 2004
- 年：2004
- 卷：43
- 期：42
- 页码：13598 - 13603
- 全文大小：88K
- 年卷期：v.43,no.42(October 26, 2004)
- ISSN：1520-4995
文摘
The purple chromoprotein (asFP595) from Anemonia sulcata belongs to the family of greenfluorescent protein (GFP). Absorption and emission spectra of asFP595 are similar to those of a numberof recently cloned GFP-like red proteins of the DsRed subfamily. The earlier proposed asFP595chromophore structure [Martynov, V. I.; et al. (2001) J. Biol. Chem. 276, 21012-21016] was postulatedto result from an "alternative cyclization" giving rise to a pyrazine-type six-membered heterocycle. Herewe 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]. NMRspectroscopic studies of four chromophore-containing peptides (chromopeptides) isolated under mildconditions from enzymatic digests of asFP595 and one chromopeptide obtained from DsRed revealedthat 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 proteolysisproducts of an isolated full-length polypeptide containing a GFP-type chromophore already formed andarrested at an earlier stage of maturation. The two other asFP595 chromopeptides were isolated as proteolysisproducts of the purified chromophore-containing C-terminal fragment. One of these has an oxo group atMet-65 C
and is a hydrolysis product of another one, with the imino group at Met-65 C. TheN-unsubstituted imino moiety of the latter is generated by spontaneous polypeptide chain cleavage at avery unexpected site, the former peptide bond between Cys-64 C' and Met-65 N. Our data stronglysuggest that both zFP538 and asFP595 could be attributed to the DsRed subfamily of GFP-like proteins.
and is a hydrolysis product of another one, with the imino group at Met-65 C. TheN-unsubstituted imino moiety of the latter is generated by spontaneous polypeptide chain cleavage at avery unexpected site, the former peptide bond between Cys-64 C' and Met-65 N. Our data stronglysuggest that both zFP538 and asFP595 could be attributed to the DsRed subfamily of GFP-like proteins.