共查询到20条相似文献,搜索用时 31 毫秒
1.
Two c-type cytochromes from Chromatium vinosum have been partially purified and characterized. Cytochrome c550, which appears to function as an electron carrier in the cyclic electron transport chain of this photosynthetic purple sulfur bacterium, has a molecular weight of approximately 15,000 and an oxidation-reduction midpoint potential (Em) of + 240 mV at pH 7.4. It has (in the reduced form) an α band at 550 nm and a β band at 520 nm. Cytochrome c551 is characterized by absorbance maxima at 354 and 409 nm in the oxidized form and 418, 523, and 551 nm in the reduced form. The reduced cytochrome reacts with CO. Cytochrome c551 is a monomeric protein with a molecular weight of 18,800 ± 700 and Em = ?299 ± 5 mV (pH independent between pH 6.3 and 8.0). It appears to lack a methionine axial ligand as indicated by the absence of an absorbance band at 695 nm in the oxidized form. 相似文献
2.
Four c-type cytochromes were purified by several procedures including chromatography on DEAE-Sepharose CL-6B, Phenyl-Sepharose CL-4B and Sephadex G-75, G-100 and G-200 and chromatofocusing. Cytochrome c-551 had a pI value of 5.2 and an Mr of 260 000 consisting of six non-covalently bound polypeptides each with an Mr of 43 000, and contained four to five haems. Cytochrome c-552.5 had a pI value of 4.8 and an Mr of 56 000 consisting of two polypeptides with the same Mr 29 000, and contained two haems. Cytochromes c-551 and c-552.5 were reduced by ascorbate to about 70 and 60% of the fully dithionite-reduced values, respectively, and both were essential components in the thiosulphate-oxidizing multi-enzyme system (other components of the system were ‘enzyme A’, ‘enzyme B’ and sulphite: cytochrome c oxidoreductase). These two cytochromes functioned as electron carriers and effectors in the oxidation of thiosulphate. Some evidence suggested that cytochrome c-551 might be a specialized electron transfer component for sulphonate-sulphur oxidation. Both cytochromes could be reduced by thiosulphate in the presence of enzymes A and B. Cytochrome c-550 (basic) and cytochrome c-550 (acidic) were small proteins with Mr 15 000 and 14 000 and pI values of over 8 and 5, respectively. Their physiological role is uncertain. 相似文献
3.
Two soluble cytochromes of the C-type, cytochrome c-551 andcytochrome c-550, were purified from the bacteriochlorophyll-containingcells of a facultative methylotroph, Protaminobacter ruber StrainNR-1, by ion-exchange chromatography and gel-filtration. Cytochrome c-551 had absorption maxima at 551, 522 and 416 nmin the reduced form, and at 525, 410 and 273 nm in the oxidizedform. This cytochrome was a slightly basic protein with an isoelectricpoint of 8.4. It had a mid-point redox potential of 272 mV atpH 7.0. The molecular weight of this protein was 13,500 and13,700 by sodium dodecylsulfate polyacrylamide gel electrophoresis(SDS-PAGE) and gel-filtration, respectively. Cytochrome c-550 had absorption maxima at 550, 522 and 415 nmin the reduced form, and at 527, 409 and 278 nm in the oxidizedform. This cytochrome was acidic, having an isoelectric pointof 4.3. It had a mid-point redox potential of 227 mV at pH 7.0.Its molecular weight was 19,500 and 22,000 by SDS-PAGE and gel-filtration,respectively. (Received August 4, 1984; Accepted October 22, 1984) 相似文献
4.
Roger C. Prince Karen E. Stokley Copper E. Haith Holger W. Jannasch 《Archives of microbiology》1988,150(2):193-196
Naturally grown Beggiatoa filaments, occurring in massive near-mono-cultures at a “black smoker” wall of the Guaymas Basin hydrothermal vent site, were harvested and used for the analysis of their cytochromes. The cytochromes have been characterized by gel permeation chromatography, optical spectroscopy and redox potentiometry. Only c-type cytochromes were detected; a small, high potential cytochrome c that seems typical of its class, and a large complex (Mr 210,000) containing at least four thermodynamically distinct c-type hemes, which was partially dissociated by chromatography on DEAE-Sepharose. The hemes of the large complex have appropriate oxidation-reduction midpoint potentials (Em7 +240 mV, +15 mV,-160 mV,-340 mV) to be involved in the metabolism of sulfide, which is presumed to be the source of reductant for this organism. 相似文献
5.
The homoacetogenic bacteria Sporomusa ovata and Sporomusa sphaeroides were grown on betaine, betaine + formate, and acetoin in the absence of carbon dioxide, and the formation of membrane-bound cytochromes was determined. In S. sphaeroides, the growth substrate had little influence on the expression of cytochromes. In contrast, membranes from betaine-or acetoin-grown S. ovata cells had an 11-or 3-fold higher cytochrome b content than cells grown on betaine + formate. The cytochrome c content was reduced below the detection level after growth on the latter two substrates. The cytochromes in the membranes of S. sphaeroides and S. ovata were characterized by low-temperature difference spectroscopy, hemochrome difference spectroscopy, and redox potentiometry. Membranes of S. ovata were shown to contain two b-type cytochromes with Em,7=-153±10 mV and Em,7=-226±14 mV and two c-type cytochromes with Em,7=-86±6 mV and Em,7=-265±10 mV. In S. sphaeroides also two b-type cytochromes with Em,7=-165±7 mV and Em,7=-241±2 mV and two c-type cytochromes with Em,7=-101±4 mV and Em, 8.5=-338±9 mV could be distinguished. Cell extracts of S. sphaeroides were shown to contain all the enzymes of the acetyl-CoA (Wood) pathway. The degradation pathways of the substrates tested and the possible role of the cytochromes are discussed.Abbreviations Em,7
midpoint potential at pH 7 and 25°C
- H4F
tetrahydrofolate 相似文献
6.
1. The cytochromes of chromatophores from photosynthetically grown Rhodopseudomonas capsulata have been characterised both spectrally, using the carotenoid free mutant Ala Pho+, and thermodynamically, using the technique of redox titrations. Five cytochromes were present; two cytochromes b, E′0 = 60 mV at pH 7.0; and three cytochromes c, E′0 = 340 mV, , E′0 = 0 mV at pH 7.0.2. Redox titrations at different values of pH indicated that the mid point potentials of all the cytochromes varied with pH over some parts of the range between pH 6 and 9, with the possible exception of cytochrome c340.3. The effects of succinate and NADH on the steady state reduction of the cytochromes are reported. Succinate could reduce cytochromes c340, c120 and b60; NADH could reduce cytochromes c340, c120, b60 and b?25. Cytochrome c0 could be reduced by dithionite but not by the other substrates tested. 相似文献
7.
D. Zannoni 《Archives of microbiology》1984,140(1):15-20
The cytochromes of photosynthetically grown Rhodopseudomonas blastica have been thermodynamically characterized using the technique of redox titrations. Six cytochromes were present; two cytochromes c, E
m7= +295mV, E
m7=+345mV; and four cytochromes b, E
m7=+290mV, E
m7=+130mV, E
m7=+60mV, E
m7=-4mV. These cytochromes were tightly bound except for cytochrome c with E
m7 of+345mV which was mostly present in the soluble cell extracts.The effects of cyanide on both the cytochrome c oxidase activity and the NADH-dependent respiration, revealed the presence of a branched respiratory chain, one branch leading to a cyanide-resistant oxidase containing pathway and the other including the cyanide-sensitive cytochrome c-oxidase.The effects of antimycin A, myxothiazol and 5-undecyl-6-hydroxy-4,7-dioxobenzothiazole (UHDBT) on the steadystate NADH-dependent respiration were also studied. Antimycin A and myxothiazol appeared to act at the level of the ubiquinol-cytochrome c oxidoreductase while UHDBT drastically affected both respiratory branches.Absorption spectra of chromatophore photopigments resulted to be similar to those reported in many species of facultative photosynthetic bacteria although carotenoid absorption maxima were blue-shifted by 5 nm.The light-induced oxygen reduction performed by chromatophores from R. blastica suggested a strict interaction between photosynthetic and respiratory apparatuses. 相似文献
8.
《FEMS microbiology letters》1986,33(1):1-8
Three acidicc-type cytochromes (c-552,c-550 andc′) were purified from the soluble fraction ofRhodopseudomonas marina. Cytochromec′ is a high-spin cytochrome capable of binding carbon monoxide reversibly to its reduced form. It occurs as a dimer with anMr of 36700 (estimated by gel filtration) while the monomer has anMr of 17800 (determined by SDS-acrylamide gel electrophoresis). Cytochromec′ has a midpoint redox potential of +73 mV and an isoelectric point at pH 4.3. Cytochromesc-550 andc-552 are typical low-spin cytochromes. Cytochromec-550 has anMr of 12500, an isoelectric point at pH 4.5 and a negative redox potential of −163 mV. The molecular properties of cytochromec-552 are as follows:Mr, 18000; isoelectric point, pH 5.4; redox potential, +283 mV. 相似文献
9.
Three c-type cytochromes isolated from Nitrobacter agilis were purified to apparent homogeneity: cytochrome c-553, cytochrome c-550 and cytochrome c-549, 554. Their amino acid composition and other properties were studied. Cytochrome c-553 was isolated as a partially reduced form and could not be oxidized by ferricyanide. The completely reduced form of the cytochrome had absorption maxima at 419, 524 and 553 nm. It had a molecular weight of 25 000 and dissociated into two polypeptides of equal size of 11 500 during SDS gel electrophoresis. The isoelectric point of cytochrome c-553 was pH 6.8. The ferricytochrome c-550 exhibited an absorption peak at 410 nm and the ferrocytochrome c showed peaks at 416, 521 and 550 nm. The molecular weight of the cytochrome estimated by gel filtration and by SDS gel electrophoresis was 12 500. It had an Em(7) value of 0.27 V and isoelectric point pH 8.51. The N-terminal sequence of cytochrome c-550 showed a clear homology with the corresponding portions of the sequences of other c-type cytochromes. Cytochrome c-549, 554 possessed atypical absorption spectra with absorption peaks at 402 nm as oxidized form and at 419, 523, 549 and 554 nm when reduced with Na2S2O4. Its molecular weight estimated by gel filtration and SDS polyacrylamide gel electrophoresis was 90 000 and 46 000, respectively. The cytochrome had an isoelectric point of pH 5.6. Cytochrome c-549, 554 was highly autoxidizable. 相似文献
10.
Periplasmic extract from Desulfovibrio desulfuricans (NCIMB 8372) was found to contain two different c-type cytochromes. One is tetraheme cytochrome c3 and the other is monoheme cytochrome c553. Cytochrome c3 could be purified by a procedure involving only one chromatographic step, whereas cytochrome c553 required several such steps. Cytochrome c3 was found to have a relative molecular mass of 14300 and an isoionic point higher than 9. Analysis of the redox potentials indicated one heme at -260 mV and three hemes around -330 mV. Cytochrome c553 had a relative molecular mass of 7200, an isoionic point higher than 9 and a redox potential of 0 mV. 相似文献
11.
1. The effects of varying the ambient oxidation/reduction potential on the redox changes of cytochromes c, cytochromes b and P605 induced by a laser flash in chromatophores from Rhodopseudomonas capsulata Ala Pho+ have been investigated.2. The appearance and attenuation of the changes with varying ambient redox potential show that, of the cytochromes present, cytochromes c with Em7 = 340 mV and 0 mV, and cytochrome b, Em7 = 60 mV were concerned with photosynthetic electron flow.3. The site of action of antimycin was shown to be between cytochrome b60 and a component, as yet unidentified, called Z.4. The appearance or attenuation of laser-induced changes of cytochromes c0 and b60 on redox titration was dependent on pH, but no effect of pH on the cytochrome c340 titration was observed.5. The dependence on ambient redox potential of the laser-induced bleaching at 605 nm enabled identification of the mid-point potentials of the primary electron donor (Em7 = 440 mV) and acceptor (Em7 = ?25 mV).6. The interrelationship of these electron carriers is discussed with respect to the pathway of cyclic electron flow. 相似文献
12.
Katsuhiko Yoshimatsu Taketomo Fujiwara Yoshihiro Fukumori 《Archives of microbiology》1995,163(6):400-406
Cytochrome c-550 was purified from Magnetospirillum magnetotacticum to an electrophoretically homogeneous state, and some of its properties were determined. The cytochrome showed absorption peaks at 528 and 409 nm in the oxidized form, and at 550, 521, and 414 nm in the reduced form. Its midpoint redox potential at pH 7.0 was determined to be +289 mV. The primary structure of cytochrome c-550 was determined. Cytochrome c is composed of 97 amino acid residues, and its molecular weight was calculated to be 10,873, including heme c. Its primary structure is very similar to those of Rhodospirillum fulvum and Rhodospirillum molischianum cytochromes c
2, suggesting that M. magnetotacticum is phylogenetically related to photosynthetic bacteria. 相似文献
13.
Two types of cytochrome cd1 in the aerobic photosynthetic bacterium, Erythrobacter sp. OCh 114 总被引:1,自引:0,他引:1
Components I and II of cytochrome cd1 which had different spectral features were purified from the aerobic photosynthetic bacterium, Erythrobacter sp. strain OCh 114. Component I showed an absorption maxima at 700 and 406 nm in the oxidized form, and at 621, 552.5, 548 and 416 nm in the reduced form. Component II showed an absorption maxima at 635 and 410 nm in the oxidized form and at 628, 552.5, 548 and 417 nm in the reduced form. The relative molecular mass, Mr, of both cytochromes was determined to be 135,000 with two identical subunits. Components I and II showed pI values of 7.6 and 6.8, respectively. The redox potential of hemes ranged from +234 mV to +242 mV, except for the heme d1 of component I (Em7 = +134 mV). Components I and II showed both cytochrome c oxidase and nitrite reductase activities. Cytochrome c oxidase activity was strongly inhibited by a low concentration of nitrite and cyanide. Erythrobacter cytochromes c-551 and c-552 were utilized as electron donors for the cytochrome c oxidase reaction. The high affinity of cytochrome c-552 to component II (Km = 1.27 microM) suggested a physiological significance for this cytochrome. Erythrobacter cytochromes cd1 are unique in their presence in cells grown under aerobic conditions as compared to other bacterial cytochromes cd1 which are formed only under denitrifying conditions. 相似文献
14.
Succinate-cytochrome c reductase can be easily solubilized in a phospholipid mixture (1:1, lysolecithin:lecithin) in the absence of detergents. The resulting solution contains two b cytochromes with half-reduction potentials of 95 ± 10 mV (b561), and 0 ± 10 mV (b566) and cytochrome c1 (Em 7.2 = +280±5 mV). The oxidation-reduction midpoint potentials obtained by optical potentiometric titrations are identical to those determined by the EPR titrations and are 40–60 mV higher than the corresponding midpoint potentials of these cytochromes in intact mitochondria. In contrast to detergent-suspended preparations, no CO-sensitive cytochrome b can be detected in the phospholipid-solubilized preparation or intact mitochondria. The half-reduction potential of cytochrome b566 is pH-dependent above pH 7.0 (?60 mV/pH unit) while that of b561 is essentially pH-independent from pH 6.7–8.5, in contrast to its pH dependence in intact mitochondria. EPR characterizations show the presence of three oxidized low-spin heme-iron signals with g values of 3.78, 3.41 and 3.37. The identification of these signals with cytochromes b566 (bT), b561 (bK) and c1 respectively is made on the basis of redox midpoint potentials. No significant amounts of oxidized high-spin heme-iron are detectable. In addition, the preparation contains four distinct types of iron-sulfur centers: S1 and S2 (Em 7.4 = ?260 mV and 0 mV), and two iron-sulfur proteins which are associated with the cytochrome b-c1 complex: Rieske's iron-sulfur protein (Em 7.4 = +280 mV) and Ohnishi's Center 5 (Em 7.4 = +35 mV). 相似文献
15.
Dithionite reduced difference spectra of extracts of denitrifying pseudomonads revealed small absorption maxima at 567 and 539 nm, suggestive of α and β bands of a new b type cytochrome. The new pigment was present in cells grown both aerobically and anaerobically and was located in the particulate fraction of extracts. These extracts also contained, in much higher concentrations, additional pigments resembling cytochromes c553 and b559, which were readily reduced by NADH or endogenous substrates, although a small proportion of the b559 required dithionite for complete reduction. In contrast, most of the new 567 pigment was not readily reduced by NADH, succinate, or endogenous substrates, and it was most easily visualized with dithionite in the sample cuvette, and either endogenous substrates or NADH in the reference cuvette. Dyes of low redox potential such as benzyl viologen (Em,7 = ?359 mV), phenosafranine (Em,7 = ?250 mV) and reduced janus green (Em,7 = ?225 mV) could substitute for dithionite as reductant for the new 567 pigment. Cresyl violet (Em,7 = ?160 mV) caused partial reduction. However, redox compounds of higher potential such as reduced indigo carmine, (Em,7 = ?125 mV) reduced methylene blue (Em,7 = ?11 mV), ferrooxalate and ascorbate could not replace dithionite as reductant. Most of the cytochrome b559 and the c553 were reduced by ascorbate. Thus the new 567 pigment appears to have a mid-point potential between ?225 and ?125 mV, well below most of the cytochrome b559. The new 567-nm pigment was rapidly oxidized by brief but vigorous aeration and was also slowly and partially re-reduced when concentrated extracts were allowed to stand without aeration. A more complete reduction of the 567 pigment was readily obtained by the addition of a mixture of NADH and FAD. The 567 pigment was observed in several denitrifying pseudomonads, P. fluorescens, P. stutzeri and also in Micrococcus denitrificans, but was not detectable in the non-denitrifiers Escherichia coli or Aerobacter aerogenes. 相似文献
16.
《FEBS letters》1986,198(1):119-124
The respiratory electron-transport chain of heterotrophically dark-grown Chloroflexus aurantiacus has been investigated. Membranes isolated from these cells have been shown to contain at least three c-type cytochromes (Em, 7.0 255,180, and 10 mV), three b-type cytochromes (Em, 7.0 of 210, 60 and −65 mV) and two cytochromes of the a type with Em, 7.0 of 330 and 190 mV. Spectroscopic evidence from CO-difference spectra, CN−-duference spectra and spectra at fixed oxidation-reduction potentials suggests that the two a-type components may be analogous to cytochromes a and a3 of mitochondria. The analyses of the effects induced by CN−, myxothiazol and antimycin A on both steady-state respiratory activities and semi-rapid oxidation-reduction kinetic patterns of c- and a-type cytochromes indicate the presence of a branched respiratory chain. Growth of Chloroflexus in medium lacking added copper diminished the concentration of the a-type cytochromes but not those of cytochromes of the b and c type. 相似文献
17.
Ralf Kömen Davide Zannoni W. John Ingledew Karin Schmidt 《Archives of microbiology》1991,155(4):382-390
The electron transport system of autotrophically grown Alcaligenes eutrophus H16 has been investigated by spectroscopic and thermodynamic approaches. The results have been interpreted as evidence that isolated membranes contain a branched respiratory chain composed of three c-type haems (E
m,7=+160 mV, + 170 mV, and + 335 mV), five b-type haems (E
m,7=+ 5 mV, + 75 mV, + 205 mV, + 300 mV, and + 405 mV), two (possibly three) a-type haems [E
m,7= + 255 mV, + 350 mV, (+ 420 mV)], and nne d-type haem. EPR-analysis of the signals at g=1.93, g=2.02, and g=1.90 revealed the presence of iron-sulphur centres diagnostic of complexes I (NADH dehydrogenase), II (succinate dehydrogenase), and III (ubiquinol/cytochrome c oxidoreductase). The low potential b haems (+ 5 mV and + 75 mV) plus the Rieske protein (g=1.90, E
m,7=+ 280 mV), thought to be part of an orthodox bc
1 complex, were present in low amounts as compared to their counterparts in membranes from Paracoccus denitrificans.CO-difference spectra in the presence of either succinate, NADH, hydrogen, ascorbate/TMPD, and/or dithionite as reductants, suggested the existance of four different oxidases composed by bo-, cb-, a-, and d-type haems.It is concluded that in contrast to other chemolithotrophes, e.g. P. denitrificans, autotrophic growth of Alcaligenes eutrophus utilizes a respiratory system in which the bc
1 complex containing pathway is only partially involved in electron transport.Abbreviations Cytochrome c-551, number
wavelength in nm
- Cytochrome c
270, number
mid-point potential in mV
-
E
m,7
mid-point potential of an oxidation-reduction couple at pH 7.0
- KP
buffer, potassium phosphate-buffer
- OD
optical density at 436 nm, 1 cm light path
- TMPD
N,N,N,N-tetramethyl-p-phenylenediamine 相似文献
18.
Extinction coefficients for cytochrome b and c1 in the isolated cytochrome bc1 complex from Rhodopseudomonas sphaeroides GA have been determined. They are 25 mM?1.cm?1 at 561 nm for cytochrome b and 17.4 mM?1.cm?1 at 553 nM for cytochrome c1 for the difference between the reduced and the oxidized state. Cytochrome b is present in two forms in the complex. One form has an Em7 of 50 mV, an α-peak of 557 nm at liquid N2 temperature and of 561 nm at RT, which is red-shifted by antimycin A. The other form has an Em7 of ?90 mV, a double α-peak of 555 and 561 nm at liquid N2 temperature corresponding to 559 and 566 nm at RT. The absorption at 566 nm is red-shifted by myxothiazol. The two shifts are independent of each other. Both midpoint potentials of cytochromes b are pH-dependent. The redox center compositions of the cytochrome bc1 complexes from Rhodopseudomonas sphaeroides and from mitochondria are identical. 相似文献
19.
D B Knaff T M Worthington C C White R Malkin 《Archives of biochemistry and biophysics》1979,192(1):158-163
Three membrane-bound cytochromes from the photosynthetic purple sulfur bacterium Chromatium vinosum have been partially separated from other membrane components by treatment with deoxycholate followed by ammonium sulfate fractionation and chromatography on Bio-Gel A1.5. Cytochrome c555, present in relatively small amounts in the deoxycholate extract, has an α-band that splits into two bands at 548 and 552.5 nm at liquid nitrogen temperature. The major components of the deoxycholate extract are cytochromes c553 and b560, which are present in essentially equimolar amounts through all the stages of the purification procedure. 相似文献
20.
A thiosulphate-cytochrome c reductase was highly purified from Chlorobium thiosulphatophilum and its properties were studied. The enzyme catalyses reduction with Na2S2O3 of c cytochromes, including cytochrome c-551 of the bacterium. Cytochrome c (555, C. thiosulphatophilum) does not react directly with the enzyme at an appreciable rate but stimulates greatly the reduction by the enzyme of cytochrome c-551 with Na2S2O3. The reduction of c cytochromes catalysed by the enzyme is strongly inhibited by cyanide and sulphite.Cytochrome c (553, C. thiosulphatophilum), a c-type cytochrome with covalently bound flavin, was found to catalyse reduction with sulphide of c cytochromes, including cytochrome c-555. The reaction is strongly inhibited by cyanide. Cyanide seems to combine strongly with cytochrome c-553 probably at the flavin moiety. Thus, the absorption spectrum attributable to flavin of the haemoprotein is changed on addition of cyanide, and neither the original spectrum nor the activity reappears even after the cyanide-treated cytochrome has been subjected to gel filtration with a Sephadex G-25 column or to isoelectric focusing. 相似文献