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1.
Curtis W. Hoganson Demetrios F. Ghanotakis Gerald T. Babcock Charles F. Yocum 《Photosynthesis research》1989,22(3):285-293
Manganese in the oxygen-evolving complex is a physiological electron donor to Photosystem II. PS II depleted of manganese may oxidize exogenous reductants including benzidine and Mn2+. Using flash photolysis with electron spin resonance detection, we examined the room-temperature reaction kinetics of these reductants with Yz
+, the tyrosine radical formed in PS II membranes under illumination. Kinetics were measured with membranes that did or did not contain the 33 kDa extrinsic polypeptide of PS II, whose presence had no effect on the reaction kinetics with either reductant. The rate of Yz
+ reduction by benzidine was a linear function of benzidine concentration. The rate of Yz
+ reduction by Mn2+ at pH 6 increased linearly at low Mn2+ concentrations and reached a maximum at the Mn2+ concentrations equal to several times the reaction center concentration. The rate was inhibited by K+, Ca2+ and Mg2+. These data are described by a model in which negative charge on the membrane causes a local increase in the cation concentration. The rate of Yz
+ reduction at pH 7.5 was biphasic with a fast 400 s phase that suggests binding of Mn2+ near Yz
+ at a site that may be one of the native manganese binding sites.Abbreviations PS II
Photosystem II
- YD
tyrosine residue in Photosystem II that gives rise to the stable Signal II EPR spectrum
- Yz
tyrosine residue in Photosystem II that mediates electron transfer between the reaction center chlorophyll and the site of water oxidation
- ESR
electron spin resonance
- DPC
diphenylcarbazide
- DCIP
dichlorophenolindophenol 相似文献
2.
Selective solubilization of Photosystem II membranes with the non-ionic detergent octyl thioglucopyranoside has allowed the isolation of a PS II system which has been depleted of the 22 and 10 kDa polypeptides but retains all three extrinsic proteins (33, 23 and 17 kDa). The PS II membranes which have been depleted of the 22 and 10 kDa species show high rates of oxygen evolution activity, external calcium is not required for activity and the manganese complex is not destroyed by exogenous reductants. When we compared this system to control PS II membranes, we observed a minor modification of the reducing side, and a conversion of the high-potential to the low-potential form of cytochrome b
559.Abbreviations Chl-
chlorophyll
- DCBQ-
2,5-dichloro-p-benzoquinone
- DCMU-
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- ESR-
electron spin resonance
- MES-
2-(N-morpholino)ethanesulfonic acid
- OTG-
octyl--d-thioglucopyranoside
- PS II-
Photosystem II
- PEG-
polyethylene glycol, Mr=6000
- Tris-
2-amino-2-hydroxyethylpropane-1,3-diol 相似文献
3.
Touloupakis E Gessmann R Kavelaki K Christofakis E Petratos K Ghanotakis DF 《The FEBS journal》2006,273(12):2684-2692
A novel, type 1 ribosome-inactivating protein designated charybdin was isolated from bulbs of Charybdis maritima agg. The protein, consisting of a single polypeptide chain with a molecular mass of 29 kDa, inhibited translation in rabbit reticulocytes with an IC50 of 27.2 nm. Plant genomic DNA extracted from the bulb was amplified by PCR between primers based on the N-terminal and C-terminal sequence of the protein from dissolved crystals. The complete mature protein sequence was derived by partial DNA sequencing and terminal protein sequencing, and was confirmed by high-resolution crystal structure analysis. The protein contains Val at position 79 instead of the conserved Tyr residue of the ribosome-inactivating proteins known to date. To our knowledge, this is the first observation of a natural substitution of a catalytic residue at the active site of a natural ribosome-inactivating protein. This substitution in the active site may be responsible for the relatively low in vitro translation inhibitory effect compared with other ribosome-inactivating proteins. Single crystals were grown in the cold room from PEG6000 solutions. Diffraction data collected to 1.6 A resolution were used to determine the protein structure by the molecular replacement method. The fold of the protein comprises two structural domains: an alpha + beta N-terminal domain (residues 4-190) and a mainly alpha-helical C-terminal domain (residues 191-257). The active site is located in the interface between the two domains and comprises residues Val79, Tyr117, Glu167 and Arg170. 相似文献
4.
Numerous transposed sequences of mitochondrial cytochrome oxidase I-II in aphids of the genus Sitobion (Hemiptera: Aphididae) 总被引:4,自引:1,他引:3
Polymerase chain reaction (PCR) products corresponding to 803 bp of the
cytochrome oxidase subunits I and II region of mitochondrial DNA (mtDNA
COI-II) were deduced to consist of multiple haplotypes in three Sitobion
species. We investigated the molecular basis of these observations. PCR
products were cloned, and six clones from one individual per species were
sequenced. In each individual, one sequence was found commonly, but also
two or three divergent sequences were seen. The divergent sequences were
shown to be nonmitochondrial by sequencing from purified mtDNA and Southern
blotting experiments. All seven nonmitochondrial clones sequenced to
completion were unique. Nonmitochondrial sequences have a high proportion
of unique sites, and very few characters are shared between
nonmitochondrial clones to the exclusion of mtDNA. From these data, we
infer that fragments of mtDNA have been transposed separately (probably
into aphid chromosomes), at a frequency only known to be equalled in
humans. The transposition phenomenon appears to occur infrequently or not
at all in closely related genera and other aphids investigated. Patterns of
nucleotide substitution in mtDNA inferred over a parsimony tree are very
different from those in transposed sequences. Compared with mtDNA,
nonmitochondrial sequences have less codon position bias, more even
exchanges between A, G, C and T, and a higher proportion of nonsynonymous
replacements. Although these data are consistent with the transposed
sequences being under less constraint than mtDNA, changes in the
nonmitochondrial sequences are not random: there remains significant
position bias, and probable excesses of synonymous replacements and of
conservative inferred amino acid replacements. We conclude that a
proportion of the inferred change in the nonmitochondrial sequences
occurred before transposition. We believe that Sitobion aphids (and other
species exhibiting mtDNA transposition) may be important for studying the
molecular evolution of mtDNA and pseudogenes. However, our data highlight
the need to establish the true evolutionary relationships between sequences
in comparative investigations.
相似文献
5.
Using the non-ionic detergent dodecyl--D-maltoside we have developed a preparative method for the isolation of the 43 kDa, 47 kDa and D1-D2-Cyt b
559 species directly from thylakoid membranes. In contrast to previous procedures the photosynthetic membrane was exposed only to one mild detergent and that resulted in more stable preparations. The isolated species were examined spectroscopically and it was found that even under these mild conditions the D1-D2-Cyt b
559 did not retain the primary quinone QA.Abbreviations PS II
Photosystem II
- CF
Coupling Factor
- LHC
Light Harvesting Complex
- MES
4-morpholine-ethanesulfonic acid
- BIS-TRIS
2-[Bis (2-hydroxyethyl) amino]-2-hydroxymethyl-propane-1,3-diol
- SMN
0.4 M sucrose/50 mM MES (pH6)/10 mM NaCl 相似文献
6.
The CP47 protein was isolated from Photosystem II membranes by using a combination of the detergents n-dodecyl-β-D-maltoside and octyl-β-D-thioglucoside. The purified CP47 was used in a series of crystallization experiments,
which yielded highly reproducible hexagonal crystals. Immunoblot analysis revealed that the isolated CP47 undergoes degradation
even under dim light conditions. This degradation takes place after the protein has been dissociated from the core complex.
Proteolysis experiments with trypsin demonstrated that the dissociation of the CP47 from the PS II core complex results in
changes that render the protein sensitive to proteolysis.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
7.
8.
Kiriakos Kotzabasis Constantina Fotinou Kalliopi A. Roubelakis-Angelakis Demetrios Ghanotakis 《Photosynthesis research》1993,38(1):83-88
The three main polyamines putrescine (Put), spermidine (Spd) and spermine (Spm) were characterized by HPLC in intact spinach leaf cells, intact chloroplasts, thylakoid membranes, Photosystem II membranes, the light-harvesting complex and the PS II complex. All contain the three polyamines in various ratios; the HPLC polyamine profiles of highly resolved PS II species (a Photosystem II core and the rection center) suggest an enrichment in the polyamine Spm.Abbreviations Chl
chlorophyll
- HPLC
high performance liquid chromatography
- LHC
light-harvesting complex
- PS II
Photosystem II
- PS II-RC
Photosystem II reaction center
- Put
putrescine
- Spd
spermidine
- Spm
spermine
- 10%S-core
D1-D2-Cyt b559-47 kD-43 kD complex 相似文献
9.
J Petersen J P Dekker N R Bowlby D F Ghanotakis C F Yocum G T Babcock 《Biochemistry》1990,29(13):3226-3231
A photosystem II complex consisting of a 47-kDa chlorophyll-binding protein (CP47), the reaction center proteins D1 and D2, and cytochrome b-559 was characterized. Trace amounts of plastoquinone were found, indicating that the primary acceptor quinone QA has been extracted during purification. However, in the presence of ferricyanide, an EPR signal with the characteristic line shape and g value of the tyrosine radicals associated with photosystem II could be photoaccumulated in the majority of the reaction centers; in the absence of ferricyanide, or under low-temperature illumination conditions, a 9.5-11-G wide signal with a Gaussian line shape was observed at g = 2.003. Neither signal is observed in D1-D2-b-559 complexes, indicating that retention of CP47 produces a more native, but quinone-depleted photosystem II reaction center. The tyrosine radical photogenerated at room temperature can be trapped at cryogenic temperatures; results are presented showing that this radical can arise from tyrosine YZ, from tyrosine YD, or from both species. Low-temperature EPR spectroscopy also revealed a pronounced split signal with contributions at g = 6.05 and g = 5.75, which is attributed to high-spin, non-heme Fe3+ with axial ligation symmetry which is probably the non-heme iron associated with the acceptor side of photosystem II. 相似文献
10.
A class of compounds, usually referred to as ADRY reagents, destabilize intermediates in the photosynthetic water-oxidizing process. The effects of these species on the reduction kinetics of Z?, the oxidized donor to P-680, have been monitored in Tris-washed chloroplasts by following the decay of EPR Signal IIf. In the presence of ADRY reagents (e.g., sodium picrate, carbonyl cyanide m-chlorophenylhydrazone) this process follows an exponential time course, the decay half-time of which decreases as the ADRY reagent concentration increases. From this pseudo-first-order behavior, the second-order rate constants for four commonly used ADRY reagents have been extracted. The ADRY-induced acceleration in Z? reduction proceeds independently of conditions imposed on the acceptor side of Photosystem II and shows no synergism with exogenous electron donors. These observations are most easily rationalized in terms of a model which proposes direct reduction of Z? by the ADRY reagent followed by regeneration of the reduced ADRY reagent in a nonspecific reaction with membrane components such as carotenoids, chlorophyll or protein. A comparison of the second-order rate constants we obtain for ADRY reagents in their reaction with Z? in Tris-washed chloroplasts with those obtained from the literature for the ADRY- reagent induced deactivation of states S2 and S3 in oxygen-evolving chloroplasts reveals a close similarity between the two processes. From this observation, a general model for the action of ADRY reagents in destabilizing the high-potential oxidizing equivalents generated in Photosystem II is proposed. 相似文献