PecE and PecF, the products of two phycoerythrocyanin lyase genes (pecE and pecF) of Mastigocladus laminosus (Fischerella), catalyze two reactions: (1) the regiospecific addition of phycocyanobilin (PCB) to Cys-alpha 84 of the phycoerythrocyanin alpha-subunit (PecA), and (2) the Delta 4-->Delta 2 isomerization of the PCB to the phycoviolobilin (PVB)-chromophore [Zhao et al. (2000) FEBS Lett. 469, 9-13]. The alpha-apoprotein (PecA) as well PecE and PecF were overexpressed from two strains of M. laminosus, with and without His-tags. The products of the spontaneous addition of PCB to PecA, and that of the reaction catalyzed by PecE/F, were characterized by their photochemistry and by absorption, fluorescence, circular dichroism of the four states obtained by irradiation with light (15-Z/E isomers of the chromophore) and/or modification of Cys-alpha 98/99 with thiol-directed reagents. The spontaneous addition leads to a 3(1)-Cys-PCB adduct, which is characteristic of allophycocyanins and phycocyanins, while the addition catalyzed by PecE and PecF leads to a 3(1)-Cys-PVB adduct which after purification was identical to alpha-PEC. The specificity and kinetics of the chromophore additions were investigated with respect to the structure of the bilin substrate: The 3-ethylidene-bilins, viz., PCB, its 18-vinyl analogue phytochromobilin, phycoerythrobilin and its dimethylester, react spontaneously to yield the conventional addition products (3-H, 3(1)-Cys), while the 3-vinyl-substituted bilins, viz., bilirubin and biliverdin, were inactive. Only phycocyanobilin and phytochromobilin are substrates to the addition-isomerization reaction catalyzed by PecE/F. The slow spontaneous addition of phycoerythrobilin is not influenced, and there is in particular no catalyzed isomerization to urobilin.
The structure of phycoviolobilin, the photoactive chromophore of K K-phycoerythrocyanin, is incompatible with a chromophore ligation to the apoprotein via SH-addition (cysteine) to a v v3,3 1 -double bond of the phycobilin. The two putative phycoerythrocyanin lyase genes of Mastigocladus laminosus, pecE and pecF, were overexpressed in Escherichia coli. Their action has been studied on the addition reaction of phycocyanobilin to apo-K K-phycoerythrocyanin (PecA). In the absence of the components of K K-PEC-phycoviolobilin lyase PecE and PecF, or in the presence of only one of them, phycocyanobilin binds covalently to PecA forming a fluorescent chromoprotein with a red-shifted absorption (V V max = 641 nm) and low photoactivity ( 6 10%). In the presence of both PecE and PecF, a chromoprotein forms which by its absorption (V V max = 565 nm) and high photoreversible photochromism (100% type I) has been identified as integral K K-phycoerythrocyanin. We conclude that PecE and PecF jointly catalyze not only the addition of phycocyanobilin to PecA, but also its isomerization to the native phycoviolobilin chromophore.z 2000 Federation of European Biochemical Societies.
With remote sensing information products becoming increasingly varied and arguably improved, scientific applications of such products rely on their quality assessment. In an operational context such as the NASA (National Aeronautics and Space Administration) information production based on the MODIS (Moderate Resolution Imaging Spectroradiometer) instrument on board Earth Observing System (EOS) Terra and Aqua satellites, efficient ways of detecting product anomaly, i.e., to discriminate between product artifacts and real changes in Earth processes being monitored, are extremely important to assist and inform the user communities about potential unreliability in the products. A technique for anomaly detection, known as MAD (the median of absolute deviate from the median), in MODIS land products via time series analysis is described, which can handle intra-and inter-annual variation in the data by using MAD statistics of the original data and their first-order difference. This method is shown to be robust and work across major land products, including NDVI, active fire, snow cover, and surface reflectance, and its applicability to multi-disciplinary products is anticipated.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.