Spectroscopic and electrochemical properties of two azurins (Az-iso1 and Az-iso2) from the obligate methylotroph Methylomonas sp. strain J and the structure of novel Az-iso2

Methylomonas sp. strain J gives rise to two azurins (Az-iso1 and Az-iso2) with methylamine dehydrogenase (MADH-Mj). The intense blue bands characteristic of Az-iso1 and Az-iso2 are observed at 621 and 616 nm in the visible absorption spectra respectively, being revealed at 620−630 nm in those of usual azurins. The EPR signal of Az-iso1, similar to usual azurins, shows axial symmetry, while the axial EPR signal of Az-iso2 involves a slightly rhombic character. The half-wave potentials (E _1/2) of the two azurins and the intermolecular electron-transfer rate constants (k _ET) from MADH-Mj to each azurin were determined by cyclic voltammetry. The E _1/2 values of Az-iso1 and Az-iso2 are +321 and +278 mV vs NHE at pH 7.0, respectively. The k _ET value of Az-iso2 is larger than that of Az-iso1 by a factor of 5. However, the electron-transfer rate of Az-iso2 is interestingly slower than those of the azurins from a denitrifying bacterium, Alcaligenes xylosoxidans NCIB 11015, and the amicyanin from a different methylotroph, Methylobacterium extorquens AM1. The structure of Az-iso2 has been determined and refined against 1.6 Å X-ray diffraction data. The whole structure of Az-iso2 is quite similar to those of azurins reported already. The Cu(II) site of Az-iso2 is a distorted trigonal bipyramidal geometry like those of other azurins, but some of the Cu-ligand distances and ligand-Cu-ligand bond angle parameters are slightly different. These findings suggest that Az-iso2 is a novel azurin and perhaps functions as an electron acceptor for MADH. Received: 23 February 1999 / Accepted: 9 September 1999     

Diffusion barriers of tripartite sporopollenin microcapsules prepared from pine pollen

Tripartite sporopollenin microcapsules prepared from pine pollen (Pinus sylvestris L. and Pinus nigra Arnold) were analysed with respect to the permeability of the different strata of the exine which surround the gametophyte and form the sacci. The sexine at the surface of the sacci is highly permeable for polymer molecules and latex particles with a diameter of up to 200 nm, whereas the nexine covering the gametophyte is impermeable for dextran molecules, with a Stokes' radius > or =4 nm (Dextran T 70), and for the tetravalent anionic dye Evans Blue (Stokes' radius = 1.3 nm). The central capsules obtained by dissolution of the sporoplasts showed strictly membrane-controlled exchange of non-electrolytes, with half-equilibration times in the range of minutes (monosaccharides, oligosaccharides) to hours (dextran molecules with Stokes' radii up to 2.5 nm). The dependence of the permeability coefficients of the nexine for non-electrolytes on Stokes' radius or molecular weight shows that the aqueous pores through the nexine are inhomogeneous with respect to their size, and that most pores are too narrow for free diffusion of sugar molecules. To explain the barrier function of the nexine for Evans Blue, it is assumed that at least the larger pores, which enable slow permeation of dextran molecules, contain negative charges.     

A novel UV laser-induced visible blue radiation from protein crystals and aggregates: scattering artifacts or fluorescence transitions of peptide electrons delocalized through hydrogen bonding?

Proteins lacking prosthetic groups and/or cofactors are known to undergo electronic excitation transitions only upon exposure to UV-C (< 280 nm) and UV-B (280-320 nm), but not UV-A (320-400 nm) photons. Here, we report the discovery of a novel excitation that peaks at approximately 340 nm and yields visible violet-blue radiation with apparent band maxima at approximately 425, 445, 470, and 500 nm. All proteins and large polypeptides examined in solid form, and in solutions, display this quenchable and photobleachable radiation which can be established not owing to aromatic sidechains. As a note of caution, we wish to state that we have not been able to completely eliminate the possibility that the radiation may be an artifact owing to second order effects such as, e.g., Raman scattering of Raman-scattered photons; however, we assert that all our experiments indicate that the radiation actually owes to some form of fluorescence. We propose that peptide electrons that have been delocalized through intramolecular or intermolecular hydrogen bond formation display these long-wavelength electronic transitions. If confirmed by future studies, this preliminary discovery may turn out to have important implications for biomolecular spectroscopy, protein crystallography, and materials science.     

Proteome modifications of blue mussel (Mytilus edulis L.) gills as an effect of water pollution

The discharge of chemicals such as oil associated or not with derived products constitutes a real threat for the environment. We report here the differential expression of the blue mussel (Mytilus edulis) gill proteins corresponding to two contaminated environmental conditions: crude oil and offshore produced water. In order to evaluate and understand contaminants, effects and adaptive response of these organisms, we identified proteins using MS. The latter can be grouped into three main classes: proteins involved in the cellular structure, in metabolism, and in defence proteins.     

Blue intensity from a tropical conifer’s annual rings for climate reconstruction: An ecophysiological perspective

We developed Blue Intensity (BI) measurements from the crossdated ring sequences of Fokienia hodginsii (of the family Cupressaceae) from central Vietnam. BI has been utilized primarily as an indirect proxy measurement of latewood (LW) density of conifers (i.e., LWBI) from high latitude, temperature-limited boreal forests. As such, BI closely approximates maximum latewood density (MXD) measurements made from soft x-ray. The less commonly used earlywood (EW) BI (EWBI) represents the minimum density of EW and is influenced by the lighter pixels from the vacuoles or lumens of cells. The correlation of our BI measurements with climate, strongest for EWBI, rivals that for total ring width (RW), and we demonstrate that it can be successfully employed as an independent predictor for reconstruction models. EWBI exhibits robust spatial correlations with winter and spring land temperature, sea surface temperature (SST) over the regional domain of ENSO, and the Standardized Precipitation Evapotranspiration Index (SPEI) over Indochina. However, in order to mitigate the effects of color changes at the heartwood – sapwood boundary we calculated ΔBI (EWBI-LWBI), and it too exhibits a significant (p < 0.05), temporally stable response to prior autumn (Oct-Nov) rainfall and winter (December to April) dry season temperature. We interpret this response as reflecting a potential cavitation defense by reducing lumen diameter as a means to safeguard hydraulic conductivity in the stem, and to prevent the xylem from imploding due to negative pressure. This study has wide implications for the further use of BI from the global tropics, though it is unclear how many tropical tree species will be appropriate for use. It seems very likely that other wood anatomical measurements can be combined with BI and RW for climate reconstruction.     

Photomorphogenic mutants of Arabidopsis thaliana reveal activities of multiple photosensory systems during light-stimulated apical-hook opening

Fluence rate-response curves were generated for red-, far-red-, and blue-light-stimulated apical-hook opening in seedlings of several photomorphogenic mutants of Arabidopsis thaliana (L.) Heynh. Compared to wild-type plants, hook opening was reduced in the phytochrome-deficient hy1, hy2, and hy6 mutants in red and far-red light at all fluence rates tested, and in low-fluence blue light, but was normal under high-irradiance blue light. In contrast, the blue-light-response mutants (blu1, blu2, and blu3) lacked the high-irradiance-dependent hook-opening response in blue light while hook opening was normal in low-fluence blue light and in red and farred light at all fluence rates tested. Hook opening in the phytochrome-B-deficient hy3 mutant was similar to wild type in all light conditions tested. The effects of the different mutations on light-induced hook opening indicate that a phytochrome(s) other than phytochrome B mediates hook opening stimulated by red, far-red and lowfluence blue light, while a blue-light-absorbing photoreceptor mediates the blue-light-sensitive high-irradiance response. Although the phytochrome and blue-light photosensory systems appear to work independently for the most part, some of their signal-transduction components may interact since the hy4, and hy5 mutants showed reduced hook-opening responses under conditions dependent on the phytochrome and blue-light-photosensory systems.We thank Jeff Young and Brian Parks for their many helpful suggestions during the progress of this research. This work was supported by National Science Foundation Grant No. DCB-9106697.     

Genomic analysis reveals widespread occurrence of new classes of copper nitrite reductases

Recently, the structure of a Cu-containing nitrite reductase (NiR) from Hyphomicrobium denitrificans (HdNiR) has been reported, establishing the existence of a new family of Cu-NiR where an additional type 1 Cu (T1Cu) containing cupredoxin domain is located at the N-terminus (Nojiri et al. in Proc. Natl. Acad. Sci. USA 104:4315-4320, 2007). HdNiR retains the well-characterised coupled T1Cu-type 2 Cu (T2Cu) core, where the T2Cu catalytic site is also built utilising ligands from neighbouring monomers. We have undertaken a genome analysis and found the wide occurrence of these NiRs, with members clustering in two groups, one showing an amino acid sequence similarity of around 80% with HdNiR, and a second group, including the NiR from the extremophile Acidothermus cellulolyticus, clustering around 50% similarity to HdNiR. This is reminiscent of the difference observed between the blue (Alcaligenes xylosoxidans) and green (Achromobacter cycloclastes and Alcaligenes faecalis) NiRs which have been extensively studied and may indicate that these also form two distinct subclasses of the new family. Genome analysis also showed the presence of Cu-NiRs with a C-terminal extension of 160-190 residues containing a class I cytochrome c domain with a characteristic beta-sheet extension. Currently no structural information exists for any member of this family. Genome analysis suggests the widespread occurrence of these novel NiRs with representatives in the alpha, beta and gamma subclasses of the Proteobacteria and in two species of the fungus Aspergillus. We selected the enzyme from Ralstonia pickettii for comparative modelling and produced a plausible structure highlighting an electron transfer mode in which the cytochrome c haem at the C-terminus can come within 16-A reach of the T1Cu centre of the T1Cu-T2Cu core.     

基于MaxEnt模型的四川王朗国家级自然保护区蓝马鸡栖息地适宜性评价

明确野生动物栖息地的空间分布格局和影响因子,是有效的栖息地管理和物种保护的基础。基于王朗国家级自然保护区自2001至2018年间的野外调查记录,首次使用MaxEnt模型对蓝马鸡（Crossoptilon auritum）栖息地适宜性进行了分析和评价。结果显示：（1）地形特征和极端时期的气候是影响蓝马鸡栖息地适宜性的主要环境因子,年最低温高于-14.4℃、海拔约2430-3100 m、坡度小于40°、靠近河流且最干月降水低于4.4 mm的区域属于蓝马鸡的适宜栖息地;（2）保护区内共有蓝马鸡栖息地102.28 km²,约占保护区总面积的三分之一,其中适宜栖息地面积59.41 km²,次适宜栖息地面积42.87 km²,为该物种提供了面积巨大且连通性极好的栖息地;（3）在不适宜蓝马鸡分布的区域,海拔过高和最干月降水量较大是限制其栖息地适宜性的主要环境因子。目前保护区内主要的人为干扰类型是牦牛和马匹的放牧活动,并且放牧强度和区域有扩大趋势,潜在威胁着蓝马鸡种群及其栖息地。因此,基于模型结果和野外调查,为避免保护区内蓝马鸡栖息地的退化和破碎化,提出了控制放牧数量和面积、增强对放牧人员的宣传教育以及定期监测河流水质的保护建议,以期促进蓝马鸡的种群和栖息地保护。