Traditional GFP-Type Cyclization and Unexpected Fragmentation Site in a Purple Chromoprotein from Anemonia sulcata, asFP595
| Autor: | Alexander S. Arseniev, Andrey Yu. Gorokhovatsky, Natalia V. Rudenko, Mikhail V Zakharov, Vasily E. Zagranichny, T. A. Balashova |
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| Rok vydání: | 2004 |
| Předmět: |
Yellow fluorescent protein
Protein Denaturation Magnetic Resonance Spectroscopy Subfamily Stereochemistry Chemical structure Green Fluorescent Proteins Peptides Cyclic Biochemistry Green fluorescent protein Chromoprotein Animals Moiety chemistry.chemical_classification biology Chemistry Hydrolysis Chromophore Anthozoa Luminescent Proteins Sea Anemones Spectrometry Fluorescence Enzyme Spectrophotometry biology.protein Peptide Hydrolases |
| Zdroj: | Biochemistry. 43:13598-13603 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi0488247 |
| Popis: | 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 C(alpha) and is a hydrolysis product of another one, with the imino group at Met-65 C(alpha). 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 N(alpha). Our data strongly suggest that both zFP538 and asFP595 could be attributed to the DsRed subfamily of GFP-like proteins. |
| Databáze: | OpenAIRE |
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