Isolation, characterization and electron microscopic single particle analysis of the NADH : ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus

The proton-translocating NADH:ubiquinone oxidoreductase (complex I) has been purified from Aquifex aeolicus, a hyperthermophilic eubacterium of known genome sequence. The purified detergent solubilized enzyme is highly active above 50 degreesC. The specific activity for electron transfer from NADH to decylubiquinone is 29 U/mg at 80 degreesC. The A. aeolicus complex I is completely sensitive to rotenone and 2-n-decyl-quinazoline-4-yl-amine. SDS polyacrylamide gel electrophoresis shows that it may contain up to 14 subunits. N-terminal amino acid sequencing of the bands indicates the presence of a stable subcomplex, which is composed of subunits E, F, and G. The isolated complex is highly stable and active in a temperature range from 50 to 90 degreesC, with a half-life of about 10 h at 80 degreesC. The activity shows a linear Arrhenius plot at 50-85 degreesC with an activation energy at 31.92 J/mol K. Single particle electron microscopy shows that the A. aeolicus complex I has the typical L-shape. However, visual inspection of averaged images reveals many more details in the external arm of the complex than has been observed for complex I from other sources. In addition, the angle (90degrees) between the cytoplasmic peripheral arm and the membrane intrinsic arm of the complex appears to be invariant.

Comparative studies on the efficiency of energy transfer of two R-phycoerythrin-C-phycocyanin conjugates constructed from sodium periodate and glutaraldehyde respectively

The C-phycocyanin and the R-phycoerythrin were purified from the blue-green alga Spirulina platensis and red alga Polysiphonia urceolata respectively. Both sodium periodate and glutaraldehyde are effective coupling agents being capable of constructing the R-phycoerythrin-C-phycocyanin conjugate, which was also called phycobiliproteins energy transfer model. The two artificial conjugates constructed with different methods were purified by Sephadex G-200 chromatography respectively. Spectra analysis indicated that energy transfer occurred in the two conjugates. The conjugate with sodium periodate had the higher efficiency of energy transfer than that with glutaraldehyde conjugate.

Isolation, properties and spatial site analysis of gamma subunits of B-phycoerythrin and R-phycoerythrin

Polysiphonia urceolata R-phycoerythrin and Porphyridium cruentum B-phycoerythrin were degraded with proteinaseK, and then the nearly native gamma subunits were isolated from the reaction mixture. The process of degradation of phycoerythrin with proteinaseK showed that the gamma subunit is located in the central cavity of (alpha beta)(6) hexamer of phycoerythrin. Comparative analysis of the spectra of the native phycoerythrin, the phycoerythrin at pH 12 and the isolated gamma subunit showed that the absorption peaks of phycoerythrobilins on alpha or beta subunit are at 535 nm (or 545 nm) and 565 nm, the fluorescence emission maximum at 580 nm; the absorption peak of phycoerythrobilins on the isolated gamma subunit is at 589 nm, the fluorescence emission peak at 620 nm which overlaps the absorption maximum of C-phycocyanin and perhaps contributes to the energy transfer with high efficiency between phycoerythrin and phycocyanin in phycobilisome; the absorption maximum of phycourobilin on the isolated gamma subunit is at 498 nm, which is the same as that in native phycoerythrin, and the fluorescence emission maximum at 575 nm.

Evolutionary analysis of phycobiliproteins: Implications for their structural and functional relationships

Phycobiliproteins, together with linker polypeptides and various chromophores, are basic building blocks of phycobilisomes, a supramolecular complex with a light-harvesting function in cyanobacteria and red algae. Previous studies suggest that the different types of phycobiliproteins and the linker polypeptides originated from the same ancestor. Here we retrieve the phycobilisome-related genes from the well-annotated and even unfinished cyanobacteria genomes and find that many sites with elevated d(N)/d(S) ratios in different phycobiliprotein lineages are located in the chromophore-binding domain and the helical hairpin domains (X and Y). Covariation analyses also reveal that these sites are significantly correlated, showing strong evidence of the functional-structural importance of interactions among these residues. The potential selective pressure driving the diversification of phycobiliproteins may be related to the phycobiliprotein-chromophore microenvironment formation and the subunits interaction. Sites and genes identified here would provide targets for further research on the structural-functional role of these residues and energy transfer through the chromophores.

The variability of eddy kinetic energy in the South China Sea deduced from satellite altimeter data

We used fifteen years (1993-2007) of altimetric data, combined from different missions (ERS-1/2, TOPEX/Poseidon, Jason-1, and Envisat), to analyze the variability of the eddy kinetic energy (EKE) in the South China Sea (SCS). We found that the EKE ranged from 64 cm(2)/s(2) to 1 390 cm(2)/s(2) with a mean value of 314 cm(2)/s(2). The highest EKE center was observed to the east of Vietnam (with a mean value of 509 cm(2)/s(2)) and the second highest EKE region was located to the southwest of Taiwan Island (with a mean value of 319 cm(2)/s(2)). We also found that the EKE structure is the consequence of the superposition of different variability components. First, interannual variability is important in the SCS. Spectral analysis of the EKE interannual signal (IA-EKE) shows that the main periodicities of the IA-EKE to the east of Vietnam, to the southwest of Taiwan Island, and in the SCS are 3.75, 1.87, and 3.75 years, respectively. It is to the south of Taiwan Island that the IA-EKE signal has the most obvious impact on EKE variability. In addition, the IA-EKE exhibit different trends in different regions. An obvious positive trend is observed along the east coast of Vietnam, while a negative trend is found to the southwest of Taiwan Island and in the east basin of Vietnam. Correlation analysis shows that the IA-EKE has an obvious negative correlation with the SSTA in Nio3 (5A degrees S-5A degrees N, 90A degrees W-150A degrees W). El Nio-Southern Oscillation (ENSO) affects the IA-EKE variability in the SCS through an atmospheric bridge-wind stress curl over the SCS. Second, the seasonal cycle is the most obvious timescale affecting EKE variability. The locations of the most remarkable EKE seasonal variabilities in the SCS are to the east of Vietnam, to the southwest of Taiwan, and to the west of Philippines. To the east of Vietnam, the seasonal cycle is the dominant mechanism controlling EKE variability, which is attributed primarily to the annual cycle there of wind stress curl. In this area, the maximum EKE is observed in autumn. To the southwest of Taiwan Island, the EKE is enlarged by the stronger SCS circulation, which is caused by the intrusion branch from the Kuroshio in winter. Finally, intra-annual and mesoscale variability, although less important than the former, cannot be neglected. The most obvious intra-annual and mesoscale variability, which may be the result of baroclinic instability of the background flow, are observed to the southwest of Taiwan Island. Sporadic events can have an important effect on EKE variability.

Response of cyanobacterial carbon concentrating system to light intensity: a simulated analysis

Cyanobacteria possess a delicate system known as the carbon concentrating mechanism (CCM), which can efficiently elevate the intracellular inorganic carbon (Ci) concentration via active transportation. The system requires energy supplied by photosystems; therefore, the activity of the Ci transporter is closely related to light intensity. However, the relationship between CCM and light intensity has rarely been evaluated. Here, we present an improved quantitative model of CCM in which light is incorporated, and developed a CCM model that modified after Fridlyand et al. in 1996. Some equations used in this model were inducted to describe the relationship between transport capacity and light intensity, by which the response of the CCM to light change is simulated. Our results indicate that the efficiency of the carbon concentrating system is sensitive to light intensity. When the external Ci concentration was low, CO2 uptake dominated the total Ci uptake with increasing light intensity, while under high external Ci concentrations HCO3- uptake primarily contributed to the total Ci uptake. Variations in the ratio of energy allocated between the transport systems could markedly affect the operation of CCM. Indeed, our simulations suggest that various combinations of Ci fluxes can provide a possible approach to detect the way by which the cell distributes energy produced by the photosystems to the two active Ci transport processes. The proportion of the energy consumed on CCM to the total energy expenditure for the fixation of one CO2 molecule was determined at 18%-40%.