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71.
J Han E T Adman T Beppu R Codd H C Freeman L L Huq T M Loehr J Sanders-Loehr 《Biochemistry》1991,30(45):10904-10913
New resonance Raman (RR) spectra at 15 K are reported for poplar (Populus nigra) and oleander (Oleander nerium) plastocyanins and for Alcaligenes faecalis pseudoazurin. The spectra are compared with those of other blue copper proteins (cupredoxins). In all cases, nine or more vibrational modes between 330 and 460 cm-1 can be assigned to a coupling of the Cu-S(Cys) stretch with Cys ligand deformations. The fact that these vibrations occur at a relatively constant set of frequencies is testimony to the highly conserved ground-state structure of the Cu-Cys moiety. Shifts of the vibrational modes by 1-3 cm-1 upon deuterium exchange can be correlated with N-H...S hydrogen bonds from the protein backbone to the sulfur of the Cys ligand. There is marked variability in the intensities of these Cys-related vibrations, such that each class of cupredoxin has its own pattern of RR intensities. For example, plastocyanins from poplar, oleander, French bean, and spinach have their most intense feature at approximately 425 cm-1; azurins show greatest intensity at approximately 410 cm-1, stellacyanin and ascorbate oxidase at approximately 385 cm-1, and nitrite reductase at approximately 360 cm-1. These variable intensity patterns are related to differences in the electronic excited-state structures. We propose that they have a basis in the protein environment of the copper-cysteinate chromophore. A further insight into the vibrational spectra is provided by the structures of the six cupredoxins for which crystallographic refinements at high resolution are available (plastocyanins from P. nigra, O. nerium, and Enteromorpha prolifera, pseudoazurin from A. faecalis, azurin from Alcaligenes denitrificans, and cucumber basic blue protein). The average of the Cu-S(Cys) bond lengths is 2.12 +/- 0.05 A. Since the observed range of bond lengths falls within the precision of the determinations, this variation is considered insignificant. The Cys ligand dihedral angles are also highly conserved. Cu-S gamma-C beta-C alpha is always near -170 degrees and S gamma-C beta-C alpha-N near 170 degrees. As a result, the Cu-S gamma bond is coplanar with the Cys side-chain atoms and part of the polypeptide backbone. The coplanarity accounts for the extensive coupling of Cu-S stretching and Cys deformation modes as seen in the RR spectrum. The conservation of this copper-cysteinate conformation in cupredoxins may indicate a favored pathway for electron transfer. 相似文献
72.
The soluble and particulate (carboxysomal) forms of ribulose 1,5-bisphosphate (RuBP) carboxylase from the cyanobacterium Chlorogloeopsis fritschii have been purified separately. A molecular weight of 520,000 was found in each case. Large (L, 53,000) and small (S, 13,000) subunits were obtained after dissociation, indicating a L8S8 quaternary structure for the enzyme from both sources. The L and S subunits are identical in molecular weight to the major polypeptides present in isolated dissociated C. fritschii polyhedral bodies (carboxysomes). Occasionally an additional polypeptide (mol. wt. 45,000) was found after dissociation of the soluble enzyme only, although the possibility that this may be due to proteolysis is not discounted. Immunochemical identity between the purified soluble and carboxysomal RuBP carboxylases was indicated by tandem-crossed and rocket immunoelectrophoresis.Abbreviations PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecyl-sulphate
- RuBP
D-ribulose 1,5-bisphosphate
- TCA
trichloroacetic acid
- LTIB
low Tris isolation buffer
- HTIB
high Tris isolation buffer
- CIE
crossed immunoelectrophoresis
- TCIE
tandem-crossed immunoelectrophoresis
- RIE
rocket immunoelectrophoresis 相似文献
73.
Effect of nutritional regime on accumulation of cobalt, manganese and zinc by green microalgae 总被引:1,自引:0,他引:1
Abstract Accumulation of cobalt, manganese and zinc by the algae Chlorella emersonii, Chlamydomonas reinhardtii and Scenedesmus obliquus has been characterized under photoautotrophic, photoheterotrophic and chemoheterotrophic nutritional regimes. All three species accumulated smaller amounts of Co2+ , Mn2+ and Zn2+ under chemoheterotrophic and photoheterotrophic conditions than under photoautotrophic conditions except in the case of cobalt accumulation by C. reinhardtii where there was little difference in the amount of cobalt accumulated under any of the nutritional regimes. Decreased accumulation of the three metals by C. emersonii and C. reinhardtii largely resulted from a decrease in the initial biosorptive phase of uptake whereas the decrease in Mn2+ and Zn2+ accumulation by C. reinhardtii under chemoheterotrophic and photoheterotrophic conditions was due to a decrease in the slow energy-dependent phase of uptake. 相似文献
74.
d-Ribulose 1,5-diphosphate carboxylase from extracts of the unicellular blue-green alga Aphanocapsa 6308 has been purified by ammonium sulphate precipitation and linear sucrose density gradient centrifugation. The molecular weight was estimated to be 525 000 and the enzyme consisted of two types of sub-unit of molecular weights 51 000 and 15 000. The small sub-units were not detected after purification involving acid precipitation but were observed if the acid precipitation step was omitted. The Michaelis constants for Mg2+ and CO2, when tested under air, were 0.35 mM and 0.071 mM respectively. Oxygen acted as a competitive inhibitor with respect to CO2, suggesting that the enzyme also acts as an oxygenase. This was confirmed by measuring ribulose diphosphate-dependent O2 uptake. A 1:1 stoichiometry between ribulose diphosphate utilization and O2 consumption was observed. 6-Phosphogluconate inhibited carboxylase activity both at high (20 mM) and low (1 mM) bicarbonate concentrations. The data are compared with the properties of ribulose diphosphate carboxylase from other autotrophic prokaryotes and from chloroplasts.Abbreviations RuDP
d-Ribulose 1,5-diphosphate
- EDTA
ethylene diamine tetraacetic acid
- GSH
reduced glutathione
- SDS
sodium dodecyl sulphate
- 6PGluc
6-phosphogluconate
- STB
supplemented Tris buffer 相似文献
75.
Joshua Denham Christopher P. Nelson Brendan J. O’Brien Scott A. Nankervis Matthew Denniff Jack T. Harvey Francine Z. Marques Veryan Codd Ewa Zukowska-Szczechowska Nilesh J. Samani Maciej Tomaszewski Fadi J. Charchar 《PloS one》2013,8(7)
Telomere length is recognized as a marker of biological age, and shorter mean leukocyte telomere length is associated with increased risk of cardiovascular disease. It is unclear whether repeated exposure to ultra-endurance aerobic exercise is beneficial or detrimental in the long-term and whether it attenuates biological aging. We quantified 67 ultra-marathon runners’ and 56 apparently healthy males’ leukocyte telomere length (T/S ratio) using real-time quantitative PCR. The ultra-marathon runners had 11% longer telomeres (T/S ratio) than controls (ultra-marathon runners: T/S ratio = 3.5±0.68, controls: T/S ratio = 3.1±0.41; β = 0.40, SE = 0.10, P = 1.4×10−4) in age-adjusted analysis. The difference remained statistically significant after adjustment for cardiovascular risk factors (P = 2.2×10−4). The magnitude of this association translates into 16.2±0.26 years difference in biological age and approximately 324–648bp difference in leukocyte telomere length between ultra-marathon runners and healthy controls. Neither traditional cardiovascular risk factors nor markers of inflammation/adhesion molecules explained the difference in leukocyte telomere length between ultra-marathon runners and controls. Taken together these data suggest that regular engagement in ultra-endurance aerobic exercise attenuates cellular aging. 相似文献
76.
Hastie CJ Borthwick EB Morrison LF Codd GA Cohen PT 《Biochimica et biophysica acta》2005,1726(2):187-193
Microcystins produced by cyanobacterial 'blooms' in reservoirs and lakes pose significant public health problems because they are highly toxic due to potent inhibition of protein serine/threonine phosphatases in the PPP family. A dehydrobutyrine (Dhb)-containing microcystin variant [Asp3, ADMAdda5, Dhb7]microcystin-HtyR isolated from Nostoc sp. was found to potently inhibit PP1, PP2A, PPP4 and PPP5 with IC50 values similar to those of microcystin-LR. However, in contrast to microcystin-LR, which forms a covalent bond with a cysteine residue in these protein phosphatases, Asp,ADMAdda,Dhb-microcystin-HtyR did not form any covalent interaction with PP2A. Since the LD50 for Asp,ADMAdda,Dhb-microcystin-HtyR was 100 microg kg(-1) compared to 50 microg kg(-1) for microcystin-LR, the data indicate that the non-covalent inhibition of protein phosphatases accounts for most of the harmful effects of microcystins in vivo. A 3-amino-6-hydroxy-2-piperidone containing cyclic peptide, nostocyclin, also isolated from Nostoc sp., was non-toxic and exhibited more than 500-fold less inhibitory potency towards PP1, PP2A, PPP4 and PPP5, consistent with the conclusion that potent inhibition of one or more these protein phosphatases underlies the toxicity of microcystins, both lacking and containing Dhb. 相似文献
77.
Cylindrospermopsin, a cyanobacterial guanidine alkaloid hepatotoxin and protein synthesis-inhibitor, was assayed for its effects on the germination of pollen from tobacco (Nicotiana tabacum cv Samsun NN). Pollen germination, measured by Alcian Blue dye-binding, was inhibited by cylindrospermopsin between 5 and 1000 microg ml(-1). As a protein synthesis-inhibitor, cylindrospermopsin did not inhibit pollen germination to the same extent as cycloheximide on a gravimetric basis, but significantly reduced the amount of (14)C-(U)-l-leucine labelling in pollen tubes. The inhibition of tobacco pollen germination may be amenable for development as a bioassay for cylindrospermopsin, although this would require a pre-concentration step for the monitoring of environmental samples. Implications of these observations for current spray-irrigation practices are discussed. 相似文献
78.
P. Keshavanath M. C. M. Beveridge † D. J. Baird L. A. Lawton ‡ A. Nimmo G. A. Codd ‡ 《Journal of fish biology》1994,45(1):123-129
Small (10 g) tilapia ( Oreochromis niloticus ) were exposed to pure and mixed populations of toxic and non-toxic strains of the cyanobacterium Microcystis aeruginosa (100% toxic, 50% toxic, 25% toxic, 0% toxic) at two particle concentrations (1 × 106 and 5 × 10s particles ml−1 ). At both concentrations there was a progressive decrease in grazing rate as the percentage of toxic cells increased. Differences in opercular beat rates, and hence the volumes of water passed over the gills, were also recorded among treatments, opercular beat rates decreasing as the percentage of toxic cells increased. Although in all treatment groups with toxic cells present, the medium had detectable levels (>250 ng I−1 ) of extracellular microcystin-LR toxin present, grazing was correlated with particle-bound rather than extracellular levels. 相似文献
79.
The purpose of this study was to evaluate the growth hormone (GH) releasing activity of orally administered His-D-Trp-Ala-Trp-D-Phe-Lys-NH2 (GHRP-6, SK&F 110679) in rats, dogs and monkeys. Rats were administered GHRP-6 orally by gavage or parenterally through femoral artery catheters. Blood was collected before and after GHRP-6 administration for estimation of plasma GH and comparison of GH changes resulting from enteral and parenteral administration of the peptide. GHRP-6 was administered to dogs intravenously (i.v.) through cephalic vein catheters, intragastrically (i.g.) through esophagostomy tubes or intraduodenally (i.d.) through vascular access ports, and blood was collected before and after peptide administration for estimation of plasma GH. Cynomolgus monkeys were administered GHRP-6 i.g., and blood was collected from abdominal aorta for estimation of changes in plasma GH. Enteral activity of GHRP-6 was observed in all 3 species tested. In rats, ED50's for enteral and parenteral administration of GHRP-6 were 4 mg/kg and 28 micrograms/kg, respectively. Thus in rats, enterally administered GHRP-6 was 0.7% as bioactive as the parenterally administered peptide. In dogs GHRP-6 was slightly less potent than in rats, with ED50's for i.g. and i.v. administration approximately 15 mg/kg and 125 micrograms/kg, respectively. However, enteral potency of GHRP-6 in dogs was 0.8% of parenteral potency, and thus, comparable to that in rats. Additionally, comparison of plasma GH levels following i.g. vs i.d. administration in dogs suggested greater activity by the i.d. route. Monkeys were the species most sensitive to enterally administered GHRP-6, with plasma GH increased in those receiving i.g. doses as low as 0.3 mg/kg and an ED50 of 0.75 mg/kg compared to 4 and 15 mg/kg in rats and dogs, respectively. The results of this study demonstrate that GHRP-6 releases GH when administered directly into the gastrointestinal tract. Although enteral activity is approximately 1% of parenteral activity, GHRP-6 is potent, especially in primates which require relatively low doses to provoke GH release. These data suggest that orally active GHRP-6 may provide a practical therapeutic alternative to parenterally administered peptides such as GHRH, especially if enteral activity is enhanced with appropriate formulation. 相似文献
80.