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1.
Jeong-Woo Choi Yun-Suk Nam Sei-Jeong Park Won-Hong Lee Dongho Kim Masamichi Fujihira 《Biosensors & bioelectronics》2001,16(9-12):819-825
Photoinduced electron transfer in the molecular electronic device consisting of protein-adsorbed hetero Langmuir–Blodgett (LB) film was investigated. Three kinds of functional molecules, cytochrome c, viologen, and green fluorescent protein (GFP) were used as an electron acceptor, a mediator, and a sensitizer, respectively. The hetero-LB film was fabricated by subsequently depositing cytochrome c and viologen onto the pretreated ITO or quartz glass. GFP adsorbed hetero-LB films were prepared by soaking the hetero-LB films into the buffer solution containing GFP. The MIM (metal/insulator/metal) structured molecular device was constructed by depositing aluminum onto the surface of the GFP-adsorbed hetero LB films. Due to the excitation by irradiation with a 460 nm monochromic light source, the photoinduced unidirectional flow of electrons in the MIM device could be achieved and was detected as photocurrents. The photoswitching function was achieved and the rectifying characteristic was observed in the molecular device. Based on the measurement of transient photocurrent of molecular device, the unidirectional flow of electrons was verified. 相似文献
2.
A bioelectronic device consisting of protein-adsorbed hetero-Langmuir-Blodgett (LB) films was investigated. Four kinds of functional molecules, cytochrome c, viologen, flavin, and ferrocene, were used as a secondary electron acceptor (A2), a first electron acceptor (A1), a sensitizer (S), and an electron donor (D), respectively. To fabricate the cytochrome c adsorbed hetero-LB film, poly-L-aspartic acid was used as the bridging molecule. The hetero-LB film was fabricated by subsequently depositing ferrocene, flavin, and viologen onto the pretreated ITO glass. Cytochrome c-adsorbed hetero-LB films were prepared by the adsorption of cytochrome c onto the poly-L-aspartic acid treated-LB films by intermolecular electrostatic attraction. Finally, the MIM (metal/insulator/metal) structured molecular device was constructed by depositing aluminum onto the surface of the cytochrome c-adsorbed hetero-LB films. Hetero-LB films were analyzed by Atomic Force Microscopy (AFM), and cytochrome c adsorption onto the films confirmed. The photoswitching function was achieved and the photoinduced unidirectional flow was in accordance with the rectifying characteristics of the molecular device. The direction of energy flow was in accordance with the energy level profile across molecular films. Based on the measurement of the transient photocurrent of the molecular device efficient directional flow of photocurrent through the redox potential difference was observed. The photodiode characteristics of the proposed bio-electronic device were verified and the proposed molecular array mimicking the photosynthetic reaction center could be usefully applied as a model system for the development of the bio-molecular photodiode. 相似文献
3.
Protein A molecular thin film was fabricated as a platform of antibody-based biosensor. For the immobilization of the protein
A thin film, a viologen multilayer was built up using the Langmuir-Blodgett (LB) technique, and then, protein A was adsorbed
on the viologen LB film by an electrostatic interaction force, which was formed as a hetero-film structure. For the deposition
of viologen, surface pressure area (π-A) isotherm was investigated. The fabricated protein A-viologen hetero LB film was investigated
using atomic force microscopy (AFM). Using the developed molecular film, antibody immobilization and fluorescence measurement
was carried out. 相似文献
4.
Ultra thin film of photosensitive polyimide having benzene and sulfonyloxyimide moieties in the main chain was prepared using a Langmuir-Blodgett (LB) technique, and then micro array pattern of the polyimide LB film on a gold substrate was obtained by deep UV lithographic technique. In order to array cytochrome c molecules along the micro-patterned gold substrate, the well-characterized monolayer of cytochrome c was immobilized with a mixed monolayer of 11-mercaptoundecanoic acid (11-MUDA) and decanethiol. The redox activity and electron transfer between cytochrome c molecular center and gold electrode interface for the self-assembled cytochrome c monolayer were investigated by cyclic voltammetry measurement. Biomolecular photodiode consisting of cytochrome c and green fluorescent protein (GFP) onto the patterned gold substrate was fabricated by self-assembly process. The integration and morphology of cytochrome c and GFP were studied from the measurements of atomic force microscopy (AFM) and fluorescence emission. Especially, current-voltage characteristics of the protein multilayers were investigated by scanning tunneling microscopy (STM) and its application in biomolecular photodiode was also examined. 相似文献
5.
Deposition behavior and photoelectrochemical characteristics of chlorophylla Langmuir-Blodgett films
Hyun-Goo Choi Byung-Keun Oh Won Hong Lee Jeong-Woo Choi 《Biotechnology and Bioprocess Engineering》2001,6(3):183-188
The deposition behavior and photoelectric response characteristics of chlorophylla (Chla) monolayers and multilayers were investigated under various film fabrication conditions. Chla LB films were deposited onto quartz and pretreated ITO glass substrates under several fabrication conditions, including surface
pressure and number of layers. The absorption spectra of Chla in a solution state and solid-like state (LB films) were fairly consistent with each other, and two absorption peaks were
found at 678 and 438 nm, respectively. The prepared Chla LB films were set into an electrochemistry cell equipped with a Pt plate as the counter electrode, and the photoelectric
response characteristics were obtained and analyzed relative to the light illumination. By considering the resulting photocurrents,
the optimal fabrication conditions for Chla LB films were determined as 20 mN/m of surface pressure and 20 layers. The action spectrum of the Chla LB films was obtained in the visible region, and was found to be in good agreement with the absorption spectrum. The possible
application of the proposed system as a constituent of an artificial color recognition device was suggested based on combining
with the photoelectric conversion property of another lightsensitive biological pigment. 相似文献
6.
Cytochrome c3 from Desulfovibrio vulgaris has four hemes per molecule, and a redox change at the hemes alters the conformation of the protein, leading to a redox-dependent change in the interaction of cytochrome c3 with redox partners (an electron acceptor or an electron donor). The redox-dependent change in this interaction was directly monitored by the high-performance electrochemical quartz crystal microbalance (EQCM) technique that has been improved to give high sensitivity in solution. In this method, cytochrome c3 molecules in solution associate electrostatically with a viologen-immobilized quartz crystal electrode as a monolayer, and redox of the associating cytochrome c3 is controlled by the immobilized viologen. This technique makes it possible to measure the access of cytochrome c3 to the electrode or repulsion from the electrode, and hence interconversion between an electrostatic complex and an electron transfer complex on the cytochrome c3 and the viologen as a mass change accompanying a potential sweep is monitored. In addition, simultaneous measurement of a mass change and a potential step reveals that the cytochrome c3 stores electrons when the four hemes are reduced (an electron pool effect), that is, the oxidized cytochrome c3 facilitates acceptance of electrons from the immobilized viologen molecule, but the reduced cytochrome c3 donates the accepted electrons to the viologen with difficulty. 相似文献
7.
The spectral changes caused by binding soft ligands to the cytochrome c iron and their correlation to ligand affinities support the hypothesis that the iron—methionine sulfur bond of this heme protein is enhanced by delocalization of the metal l2, electrons into the empty 3d orbitals of the ligand atom. These findings also explain the unique spectrum of cytochrome c in the far red. 相似文献
8.
Rui M. Almeida Sofia R. Pauleta Isabel Moura Jos J.G. Moura 《Journal of inorganic biochemistry》2009,103(9):1245-1253
The paramagnetic effect due to the presence of a metal center with unpaired electrons is no longer considered a hindrance in protein NMR spectroscopy. In the present work, the paramagnetic effect due to the presence of a metal center with unpaired electrons was used to map the interface of an electron transfer complex. Desulfovibrio gigas cytochrome c3 was chosen as target to study the effect of the paramagnetic probe, Fe-rubredoxin, which produced specific line broadening in the heme IV methyl resonances M21 and M181. The rubredoxin binding surface in the complex with cytochrome c3 was identified in a heteronuclear 2D NMR titration. The identified heme methyls on cytochrome c3 are involved in the binding interface of the complex, a result that is in agreement with the predicted complexes obtained by restrained molecular docking, which shows a cluster of possible solutions near heme IV. The use of a paramagnetic probe in 1HNMR titration and the mapping of the complex interface, in combination with a molecular simulation algorithm proved to be a valuable strategy to study electron transfer complexes involving non-heme iron proteins and cytochromes. 相似文献
9.
Interaction between uranium and the cytochrome c 3 of Desulfovibrio desulfuricans strain G20 总被引:1,自引:0,他引:1
Rayford B. Payne Laurence Casalot Tessa Rivere Jeffrey H. Terry Lise Larsen Barbara J. Giles Judy D. Wall 《Archives of microbiology》2004,181(6):451-406
Cytochrome c3 of Desulfovibrio desulfuricans strain G20 is an electron carrier for uranium (VI) reduction. When D. desulfuricans G20 was grown in medium containing a non-lethal concentration of uranyl acetate (1 mM), the rate at which the cells reduced U(VI) was decreased compared to cells grown in the absence of uranium. Western analysis did not detect cytochrome c3 in periplasmic extracts from cells grown in the presence of uranium. The expression of this predominant tetraheme cytochrome was not detectably altered by uranium during growth of the cells as monitored through a translational fusion of the gene encoding cytochrome c3 (cycA) to lacZ. Instead, cytochrome c3 protein was found tightly associated with insoluble U(IV), uraninite, after the periplasmic contents of cells were harvested by a pH shift. The association of cytochrome c3 with U(IV) was interpreted to be non-specific, since pure cytochrome c3 adsorbed to other insoluble metal oxides, including cupric oxide (CuO), ferric oxide (Fe2O3), and commercially available U(IV) oxide.An erratum to this article can be found at 相似文献
10.
The triphasic course previously reported for the reduction of cytochrome b in the succinate-cytochrome c reductase by either succinate or duroquinol has been shown to be dependent on the redox state of the enzyme preparation. Prior reduction with increasing concentrations of ascorbate leads to partial reduction of cytochrome c1, and a gradual decrease in the magnitude of the oxidation phase of cytochrome b. At an ascorbate concentration sufficient to reduce cytochrome c1 almost completely, the reduction of cytochrome b by either succinate or duroquinol becomes monophasic. Owing to the presence of a trace amount of cytochrome oxidase in the reductase preparation employed, the addition of cytochrome c makes electron flow from substrate to oxygen possible. Under such circumstances, the addition of a limited amount of either succinate or duroquinol leads to a multiphasic reduction and oxidation of cytochrome b. After the initial three phases as described previously, cytochrome b becomes oxidized before cytochrome c1 when the limited amount of added substrate is being used up. However, at the end of the reaction when cytochrome ca is being rapidly oxidized, cytochrome b becomes again reduced. The above observations support a cyclic scheme of electron flow in which the reduction of cytochrome b proceeds by two different routes and its oxidation controlled by the redox state of a component of the respiratory chain. 相似文献
11.
The photosynthetic reaction centers (RCs) classified as the group II possess a peripheral cytochrome (Cyt) subunit, which serves as the electron mediator to the special-pair. In the cycle of the photosynthetic electron transfer reactions, the Cyt subunit accepts electrons from soluble electron carrier proteins, and re-reduces the photo-oxidized special-pair of the bacteriochlorophyll. Physiologically, high-potential cytochromes such as the cytochrome c2 and the high-potential iron–sulfur protein (HiPIP) function as the electron donors to the Cyt subunit. Most of the Cyt subunits possess four heme c groups, and it was unclear which heme group first accepts the electron from the electron donor. The most distal heme to the special-pair, the heme-1, has a lower redox potential than the electron donors, which makes it difficult to understand the electron transfer mechanism mediated by the Cyt subunit. Extensive mutagenesis combined with kinetic studies has made a great contribution to our understanding of the molecular interaction mechanisms, and has demonstrated the importance of the region close to the heme-1 in the electron transfer. Moreover, crystallographic studies have elucidated two high-resolution three-dimensional structures for the RCs containing the Cyt subunit, the Blastochloris viridis and Thermochromatium tepidum RCs, as well as the structures of their electron donors. An examination of the structural data also suggested that the binding sites for both the cytochrome c2 and the HiPIP are located adjacent to the solvent-accessible edge of the heme-1. In addition, it is also indicated by the structural and biochemical data that the cytochrome c2 and the HiPIP dock with the Cyt subunit by different mechanisms although the two electron donors utilize the same region for the interactions; cytochrome c2 is recognized through electrostatic interactions while hydrophobic interactions are important in the HiPIP docking. 相似文献
12.
The reaction between membrane-bound cytochrome c and the reaction center bacteriochlorophyll g dimer P798 was studied in the whole cells and isolated membranes of Heliobacterium gestii. In the whole cells, the flash-oxidized P798+ was rereduced in multiple exponential phases with half times (t
1/2s) of 10 s, 300 s and 4 ms in relative amplitudes of 40, 35 and 25%, respectively. The faster two phases were in parallel with the oxidation of cytochrome c. In isolated membranes, a significantly slow oxidation of the membrane-bound cytochrome c was detected with t
1/2 = 3 ms. This slow rate, however, again became faster with the addition of Mg2+. The rate showed a high temperature dependency giving apparent activation energies of 88.2 and 58.9 kJ/mol in the whole cells and isolated membranes, respectively. Therefore, membrane-bound cytochrome c donates electrons to the P798+ in a collisional reaction mode like the reaction of water-soluble proteins. The rereduction of the oxidized cytochrome c was suppressed by the addition of stigmatellin both in the whole cells and isolated membranes. This indicates that the electron transfer from the cytochrome bc complex to the photooxidized P798+ is mediated by the membrane-bound cytochrome c. The multiple flash excitation study showed that 2–3 hemes c were connected to the P798. By the heme staining after the SDS-PAGE analysis of the membraneous proteins, two cytochromes c were detected on the gel indicating apparent molecular masses of 17 and 30 kDa, respectively. The situation resembles the case in green sulfur bacteria, that is, the membrane-bound cyotochrome c
z couples electron transfer between the cytochrome bc complex and the P840 reaction center complex.This revised version was published online in October 2005 with corrections to the Cover Date. 相似文献
13.
Plastoquinone as a common link between photosynthesis and respiration in a blue-green alga 总被引:3,自引:0,他引:3
The role of plastoquinone in a thermophilic blue-green alga, Shynechococcus sp., was studied by measuring reduction kinetics of cytochrome 553 which was oxidized with red flash preferentially exciting photosystem I. Sensitivity of the cytochrome reduction to DBMIB indicates that cytochrome 553 accepts electrons from reduced plastoquinone. Plastoquinone is in turn reduced in cells without electrons from photosystem II, since DCMU, which inhibited methyl viologen photoreduction more strongly than DBMIB, failed to affect the cytochrome reduction. Participation of cyclic electron transport around photosystem I in cytochrome reduction in the presence of DCMU was excluded, because methyl viologen and antimycin A had no effect on the cytochrome kinetics. On the other hand, electron donation from endogenous substrates to plastoquinone was suggested from decreases in rate of the cytochrome reduction by dark starvation of cells and also from restoration of fast reduction kinetics by the addition of exogenous substrates to or by reillumination of starved cells.KCN, which completely suppressed respiratory O2-uptake, induced a marked acceleration of the cytochrome reduction in starved cells. The poison was less or not effective in stimulating the cytochrome reduction in more extensively starved or reilluminated cells.Results indicate that plastoquinone is functioning not only in the photosynthetic but also in the respiratory electron transport chain, thereby forming a common link between the two energy conservation systems of the blue-green alga.
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14.
I. Maťchová I. Kučera O. Janiczek R. J. M. van Spanning L. F. Oltmann 《Archives of microbiology》1993,159(3):272-275
Exposure of aerobically-grown wild-type cells of Paracoccus denitrificans to a decreased aeration caused parallel increases in both PMS/ascorbate and succinate-linked activities of nitrite reductase. By contrast, the expression of the succinate-linked activity was considerably delayed in an insertion mutant specifically lacking the periplasmic 15 kDa cytochrome c-550. In this case the observed activity followed very closely the content of a 40 kDa cytochrome c. A subcellular fraction enriched in a haemoprotein of a similar apparent molecular weight showed the activity of cytochrome c peroxidase and was able to restore the antimycin-sensitive electron transport from membrane vesicles to nitrite reductase. It is concluded that P. denitrificans possesses an alternative nitrite-reducing pathway involving the 40 kDa cytochrome c instead of cytochrome c-550. This pathway branches from the respiratory chain after the cytochrome bc
1 segment.Abbreviations PAGE
polyacrylamide gel electrophoresis
- PMS
phenazine methosulphate 相似文献
15.
Chromatium vinosum cells form a vesicular type intracytoplasmic membrane system during phototrophic growth on thiosulfate.—An enzyme protein transferring electrons from thiosulfate to cytochromes of type c was enriched from S-144. The colorless thiosulfate: cytochrome c oxidoreductase was characterized by a molecular weight of 36,000 (after dodecylsulfate treatment) and 35,000 (by gel filtration). Isoelectric focusing revealed a pI range of 4.4 to 4.7. Apparent K
m values for the cytochromes tested were in the M range. — The endogenous electron acceptor compound, isolated from the chromatophore fraction P-144, was found to be a membrane-bound cytochrome c-552. The homogeneous cytochrome protein had an average pI value of 4.65 and a molecular weight of 71,500 determined by gel filtration. By dodecylsulfate electrophoresis it was cleaved into two proteins representing particle weights of 45,000 and 20,000.Abbreviations HiPIP
high potential nonheme iron protein
- IEF
isoelectric focusing
- SDS
dodecylsulfate, sodium salt
- Temed
N,N,N,N-tetramethylethylenediamine 相似文献
16.
Jeong-Woo Choi Young-Kee Kim In-Ho Lee Junhong Min Won Hong Lee 《Biosensors & bioelectronics》2001,16(9-12):937-943
The fiber-optic biosensor consisting of an acetylcholinesterase (AChE)-immobilized Langmuir–Blodegtt (LB) film was developed to detect organophosphorus compounds in contaminated water. The sensing scheme was based on the decrease of yellow product, o-nitrophenol, from a colorless substrate, o-nitrophenyl acetate, due to the inhibition by organophosphorus compounds on AChE. Absorbance change of the product as the output of enzyme reaction was detected and the light was guided through the optical fibers. The enzyme portion of the sensor system was fabricated by the LB technique for formation of the enzyme film. AChE-immobilized LB film was formed by adsorbing the enzyme molecules onto a viologen monolayer using the electrostatic force. The proposed kinetics for irreversible inhibition of organophosphorus compounds on AChE agreed well with the experimental data. The surface topography of AChE-immobilized LB film was investigated by atomic force microscope (AFM). The immobilized AChE had the maximum activity at pH 7. The proposed biosensor could successfully detect the organophosphorus compounds upto 2 ppm and the response time to steady signal of the sensor was about 10 min. 相似文献
17.
Jayanti Pande Kathleen Kinnally K. K. Thallum Balbir C. Verma Yash P. Myer Lucia Rechsteiner Hans Rudolf Bosshard 《Journal of Protein Chemistry》1987,6(4):295-319
Spectroscopically, the modification of horse heart ferricytochrome c with N-chloro-4-toluolsul-fonamide (Chloramine-T, CT) occurs through a two-step process, the disruption of the methionine-80 sulfur-iron linkage and a reagent-independent change, an intramolecular rearrangement. Chromatographic purification of the preparation at a 2.5:1 reagent-to-protein ratio, pH 8.0–8.5, yields two major products, the FII and FIII CT-cytochromes c. Both products contain modification of only the methionines, 80 and 65, to sulfoxides; both are monomeric, reduced by ascorbate, and the ferrous forms are oxidized by molecular oxygen and bind carbon monoxide. The redox potentials of FII and FIII are 135 and 175±15 mV. The FIII is indistinguishable from the native protein in its binding and the electron donor property toward mammalian cytochrome c oxidase. It also binds nearly as effectively as the native protein to yeast cytochrome c peroxidase, but is a less efficient donor. It is, however, a poor electron acceptor from both mammalian cytochrome c reductase and chicken liver sulfite oxidase. FII lacks cytochrome c oxidase activity and is also a poorer substrate for the other three enzymes. Both the derivatives are consistently better electron donors than acceptors. It is concluded that the binding of cytochrome c to cytochrome c oxidase and to cytochrome c peroxidase does not require the integrity of the methionine-80 sulfur linkage and that the complexation process has a finite degree of freedom with regard to the state of the heme crevice opening. The alterations of the oxidoreduction function have been analyzed in light of both prevailing models of cytochrome c function, the two-site model (one site for oxidizing and the other for reducing enzymes) and the single-site model (the same site for the oxidizing and reducing enzymes). These observations can be accommodated by either model, given the latitude that the binding domains for the oxidizing and the reducing enzymes have finite overlapping and nonoverlapping regions.To whom all correspondence related to the functional studies with cytochrome c peroxidase and sulfite oxidase is to be directed. 相似文献
18.
Cyanobacterial thylakoids catalyze both photosynthetic and respiratory activities. In a photosystem I-less Synechocystis sp. PCC 6803 strain, electrons generated by photosystem II appear to be utilized by cytochrome oxidase. To identify the lumenal electron carriers (plastocyanin and/or cytochromes c
553, c
550, and possibly c
M) that are involved in transfer of photosystem II-generated electrons to the terminal oxidase, deletion constructs for genes coding for these components were introduced into a photosystem I-less Synechocystis sp. PCC 6803 strain, and electron flow out of photosystem II was monitored in resulting strains through chlorophyll fluorescence yields. Loss of cytochrome c
553 or plastocyanin, but not of cytochrome c
550, decreased the rate of electron flow out of photosystem II. Surprisingly, cytochrome c
M could not be deleted in a photosystem I-less background strain, and also a double-deletion mutant lacking both plastocyanin and cytochromec
553 could not be obtained. Cytochrome c
M has some homology with the cytochrome c-binding regions of the cytochromecaa3 -type cytochrome oxidase from Bacillus spp. and Thermus thermophilus. We suggest that cytochrome c
M is a component of cytochrome oxidase in cyanobacteria that serves as redox intermediate between soluble electron carriers and the cytochromeaa3 complex, and that either plastocyanin or cytochrome c
553 can shuttle electrons from the cytochrome b6f complex to cytochrome c
M. 相似文献
19.
《Redox report : communications in free radical research》2013,18(1-2):35-41
AbstractThe polyphenolic structure common to flavonoids enables them to donate electrons and exert anti-oxidant activity. Since the mitochondrial electron transport chain consists of a series of redox inter-mediates, the effect of flavonoids in a complex mixture of polyphenols, as well as related pure flavonoids, was evaluated on the rat liver mitochondrial electron transport chain. A French maritime pine bark extract (PBE), a complex mixture of polyphenols and related pure flavonoids, was able to reduce cytochrome c reversibly, possibly by donation of electrons to the iron of the heme group; the donated electrons can be utilized by cytochrome c oxidase. Among single flavonoids tested, (-)-epicatechin gallate had the greatest ability to reduce cytochrome c. In addition, PBE competitively inhibited electron chain activity in both whole mitochondria and submitochondrial particles. A 3.5-fold increase in the apparent Km value for succinate was calculated from reciprocal plots. Among the flavonoids tested, taxifolin and (-)-epicatechin gallate showed minor inhibitory effects, while (±)-catechin and (+)-epicatechin were ineffective. Activities of NADH-ubiquinone, succinate-ubiquinone, and ubiquinol-cytochrome c reductases were inhibited by low concentrations of PBE to a similar extent. However, inhibition of cytochrome c oxidase activity required 4-fold higher PBE concen-trations. These results suggest that flavonoids reduce cytochrome c and that PBE inhibits electron transport chain activity mainly through NADH-ubiquinone, succinate-ubiquinone, and ubiquinol-cytochrome c reductases. 相似文献