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1.
Subunits (, ß, ) and mixtures of subunits ( ß, , ß , ß ) were isolated without denaturationfrom a chloroform extract of chloroplast coupling factor 1 (CF1)from maize (Zea mays var. Ushiku 5-4) and from spinach by fastprotein liquid chromatography (FPLC), on an anion-exchange columnof Mono-Q in the presence of n-octylglucoside (OG) and on achromatofocusing column of Mono-P. The ß -subunitcomplex (CF1 ß ) was the minimum unit required forATPase activity, as was confirmed by the reconstituted complexof ß and subunits. An subunit isolated from maizeinhibited the ATPase activity of CF1 ß from bothmaize and spinach. CF1 ß was found to contain anOG-dependent Mg2+-ATPase. The ATPase activity of CF1 ß required divalent cations, such as Mg2+ or Mn2+, for its expressionin the presence of OG; its optimum pH was 8.0 and it was markedlyinhibited by NaN3. The enzyme hydrolyzed ATP in prefernece toGTP but not CTP, UTP, ADP, AMP or pNPP. Lineweaver-Burk plotsof its activity were curvilinear in the range of 0.6–0.7mM ATP.Mg2+. 1Present address: Department of Biology, School of Education,Waseda University, Shinjuku-ku, Tokyo, 160 Japan. (Received February 15, 1989; Accepted April 20, 1989)  相似文献   

2.
A Ras-related NTP-binding protein was partially purified froma membrane fraction derived from the mycelia of Neurospora crassa.[-32P]ATP and [-32P]GTP were incubated with mem brane and solublefractions which were then irradiated with UV light to inducecrosslinking of tightly bound nucleotides. After SDS-polyacrylamidegel electrophoresis, blotting onto a nitrocellulose filter andautoradiography it was apparent that most of the proteins thatbound [-32P]-GTP also bound [-32P]ATP. Pretreatment of the membranefraction with Ras-specific antibody effectively blocked thebinding of [-32P]ATP and [-32P]GTP to several ATP-GTP-bindingproteins. The band of a protein with a molecular weight of 26kDa on the SDS-polyacrylamide gel cross-reacted strongly withthe Ras-specific antibody. The protein was extracted from thegel and further purified by repeated gel electrophoresis. Thepurified protein bound [-32P]ATP, [-32P]-GTP, [-32P]CTP and[-32P]UTP at 1.6x10 M and was autophosphorylated in thepresence of [-32P]ATP and [-32P]GTP at 1.7x10 M. Pretreatmentof the protein with Ras-specific antibody partially blockedthe autophosphorylation in the presence of these nucleotides.The binding of [-32P]ATP to the NTP-binding protein was blockedby addition of ATP at 10–4–10–3 M. ATP ata concentration of 10–4 M prevented the binding of [-32P]to a greater extent than did GTP at the same concentration.Binding of [-32P]CTP and [-32P]UTP to the protein was also observed. (Received October 7, 1991; Accepted July 14, 1992)  相似文献   

3.
Enzymatic Degradation of Chlorophyll in Chenopodium album   总被引:3,自引:0,他引:3  
The breakdown of chlorophyll (Chi) in crude extracts of Chenopodiumalbum (white goose foot) in the dark was examined. Derivativesof pheophorbide were formed when Chi or chlorophyllide wasincubated with depigmented crude extracts. The formation ofpheophorbide was completely prevented by heat treatment of extracts,indicating that the reaction was enzymatic, and the presenceof a Mg-releasing enzyme, the so called Mg-dechelatase, waspostulated. This hypothesis was strongly supported by the observationthat the formation of pheophorbide was inhibited by 51% by 10mM MgCl2. Analysis by high-performance thin-layer chromatography(HPTLC) and liquid chromatography (HPLC) showed that the appearanceof chlorophyllide , pheophorbide 132-hydroxychlorophyllide and pyropheophorbide was accompanied by a concomitant decreasein levels of Chi The formation of 132-hydroxychloro-phyllide was not clearly an enzymatic reaction and requires furtherexamination. It appears that Chl is degraded in a crude extractof C. album via the following enzymatically catalyzed reactions (Received September 10, 1990; Accepted November 15, 1990)  相似文献   

4.
The sexual agglutinability of haploid cells of heterothallicSaccharomyces cerevisiae was repressed when they were culturedin the absence of easily fermentable sugars, such as glucoseand mannose. The repression was reversed by the action of hormone-likesubstances of the opposite mating types. The substance producedby mating type cells was identical to subtsance-I which isknown to induce sexual agglutinability of inducible matingtype cells. The mating type cells produce a new hormone-likesubstance which induces or enhances sexual agglutinability of mating type cells. A crude fraction of the mating type-specific substance ( substance-I)was obtained by passing the culture filtrate of mating typecells through Amberlite CG-50 (H+ form), followed by elutionwith 1.5 M ammonia. 2 On leave from Osaka City University. (Received December 25, 1975; )  相似文献   

5.
The effect of growth temperature on the compositions of phospholipidsand fatty acids of the psychrotrophic bacterium Pseudomonassp. strain E-3 was studied. The composition of phospholipids(phosphatidylethanolamine, phosphatidylglycerol and cardiolipin)did not differ significantly in cells grown at 5?, 15?, and30?C. Phosphatidylethanolamine (PE) was the most abundant, amountingto over 70% of total phospholipids. When the growth temperaturewas lowered, palmitoleic [16:1(9)] and cis-vaccenic [18:1(11)]acids increased at the expense of palmitic (16:0) acid, especiallyin PE. In vivo experiments using [l-14C]16:0 showed that incorporationof radioactivity into 16:1(9) was oxygen-dependent and cerulenin-insensitive,and incorporation into 18:1(11) was cerulenin-sensitive. Theseresults suggest that 16:0 was aerobically desaturated to 16:1(9)and subsequently elongated to 18:1(11). Desaturation activityof 16:0 in the cells grown at 25?C was enhanced by loweringthe assay temperature. (Received February 12, 1987; Accepted July 10, 1987)  相似文献   

6.
Nucleoside triphosphate(NTP)-binding proteins were detectedin the crude extract of mycelia of Neurospora crassa, whichwas treated with 1% Lubrol PX and fractionated by gel filtration.Protein fractions showing the capacity to bind [35S]ATPS or[35S]GTPS were designated as AGN1 to 6. The binding of [35S]ATPSor [35S]GTPS was prevented in the presence of 0.1 mM ATP orGTP except that in fractions AGN1 and 2, the presence of GTPstimulated the binding of [35S] ATPS to ATP(NTP)-binding proteins.ATP or GTP was 1 to 2 orders of magnitude more effective thanCTP or UTP in preventing the binding of [35S]GTPS in AGN1, 2and 5. Among these fractions AGN1, 2, 5 and 6 showed activityto hydrolyze 1 nM [–32P]ATP or [–32P]GTP. NTP-bindingproteins bound with [35S]ATPS or [35S]GTPS had lower apparentmolecular weights than the same proteins without bound nucleotide.Proteins bound with [35S]ATPS or [35S]GTPS and those [32P]ADP-ribosylatedby endogenous ADP-ribosyl transferase in each fraction wereanalyzed by SDS-PAGE. About 20 species of ATP or ATP-GTP-bindingproteins were detected, several of which were ADP-ribosylated.The binding of [35S]ATPS or [35S]GTPS to NTP-binding proteinswas confirmed by the comparison of non-boiled and boiled samplesimmediately before loading to SDS-PAGE. ATP, GTP, CTP or UTPat the concentration of 0.1 mM effectively removed [33S]ATPSor [35S]GTPS bound to NTP-binding proteins. (Received December 10, 1990; Accepted April 18, 1991)  相似文献   

7.
Sixteen legumes were grown in N-free media so that N was suppliedentirely by symbiotic N2 fixation. The plant tissues were analyzedfor natural 15N abundance (expressed as 15N per mil relativeto air N2) with a ratio mass spectrometer. The nodules of desmodium,centro, siratro, soybean and winged bean showed high enrichmentin 15N (+9), while red clover showed slight enrichment (+2).The nodules of 9 other forage legumes (Townsville stylo, whiteclover, alsike clover, common vetch, Chinese milk vetch, senna,alfalfa, ladino clover, and hairy vetch) showed little enrichmentin 15N. In all the legumes investigated, particularly in the ureide-transportingplants such as desmodium, centro, siratro, soybean, winged beanand field bean, the 15N value of the shoots was negative (–3.2).The 15N value of the shoots in winged bean and field bean variedby about 1 depending on the Rhizobium strains used. The isotopicmass balance of 13 legumes indicated that isotopic fractionationoccurs during N2 fixation by the legume-rhizobia symbiosis witha preference for 14N over 15N, resulting in a 15N value of –0.2to –2 in the whole plant. The results indicate that 15N/14N isotopic discrimination witha preference for the lighter atom may occur in both N2 fixationand export of fixed N from nodules. 1Present address: Department of Soils and Fertilizers, NationalAgriculture Research Center, Kannondai, Tsukuba, Ibaraki 305,Japan. (Received October 8, 1985; Accepted April 7, 1986)  相似文献   

8.
The euryhaline charophyte Lamprothamnium papulosum (Wallr.)J. Gr. was adapted to media with decreasing salinities rangingfrom 550 to 0 mosmol kg–1. Vegetative plants grown inmedia with osmotic pressures (0) in the range of 550 to 130mosmol kg–1 maintained a constant turgor pressure () at309 + 7 mosmol kg–1. The ions K+, Na+ and Cl–, werethe predominant solutes in the vacuole. Changes in their concentrationsaccount for the variation in internal osmotic pressure (1) with,0. The divalent ions Mg2+, Ca2+ and were also present in significant amounts, but their concentrationsdid not alter with changes in, 0. In cells subjected to hypo-osmotic shock the regulation of was incomplete. The turgor pressure increased from 302 to 383mosmol kg–1. The first rapid response to the sudden decreasein 0 was a loss of K+ and Cl. In contrast to the decreasein ionic concentrations an accumulation of sucrose occurredwhich could account for the increase of . The increase in sucroseconcentration started 24 to 48 h after the downshock and reachedits highest value after 3 to 4 weeks. The sucrose concentrationin the vacuole was up to 320 mol m–3. During this timethe ionic content continued to decrease but did not counterbalancethe sucrose concentration sufficiently to regain the original. High sucrose levels accompanied by an enhanced were also observedduring the period of fructification (sexual reproduction: formationof antheridia and oogonia) in Lamprothamnium kept under conditionsof constant salinity. It is concluded that high sucrose content and elevated arecharacteristic of sexual reproduction in this charophyte. Lamprothamniumis able to tolerate different during various developmentalstages (e.g. vegetative and reproductive phases). Key words: Lamprothamnium papulosum, sucrose, turgor pressure  相似文献   

9.
Various Cucurbita seed globulins showed patterns similar toone another on SDS-gel electrophoresis, and ß bandsfor unreduced globulins and , ', and ' bands for reduced ones.On gel electrophoresis in 6 M urea, reduced globulin gave twoacidic and two basic bands. These corresponded to and ' chainsand 1 and 2 chains, respectively, identified by two-dimensionalurea-SDS gel electrophoresis. The compositions of the and ßsubunits were proposed. (Received September 8, 1977; )  相似文献   

10.
Effects of l, N6-ethenoadenylates (e-adenylates) were testedon phosphorylation, and electron transport under phosphorylation,arsenylation and quasi-arsenylation (stimulation of electrontransport in the presence of ATP, AMP and arsenate) conditionsin isolated spinach chloroplasts. -ATP as well as ATP partially inhibited ferricyanide reductionthrough binding to the chloroplast coupling factor 1 with anapparent dissociation constant (KDapp) of around 5µM,which was remarkably larger than that for ATP (ca. 2µM).e-ATP at below 500 µM had no effect on phosphorylationbut inhibited quasi-arsenylation in competition with ATP withan apparent inhibition constant (K1app) of around 60 µM. -ADP as well as ADP partially inhibited ferricyanide reductionwith a KDapp value close to that for -ATP. -ADP was phosphorylated(the apparent Michaelis constant, Kmapp=80µM) accompanyingstimulation of ferricyanide reduction to the magnitude predicted(P/e=l). -ADP-arsenylation was also detected by stimulationof ferricyanide reduction. -AMP alone caused little inhibition of ferricyanide reductionas AMP, but competitively depressed the electron transport inhibitionby ADP and ATP with a K1app value of around 200 µM. -AMPwas not effective for ADP phosphorylation but inhibited stimulationdue to quasi-arsenylation coupling in competition with AMP K1app=150µM Among the possible combinations of adenylates and -adenylatesfor quasiarsenylation, only [ATP+AMP] could couple with theenergy transduction mechanism. Based on the specificity of binding sites to adenylates and-adenylates, an attempt was made to distinguish at least four(two pairs) kinds of binding sites (at least six sites in toto)on the chloroplast coupling factor 1 for photosynthetic energytransduction. When one pair of sites is occupied by the designatedadenylates or -adenylates (allosteric effectors), the couplingfactor is thought to be in a conformation for coupling withthe energy transduction mechanism in the presence of phosphateor arsenate. 1Presented to the 1st Symposium of Japan Bioenergetics Group,December 19, 1975, Osaka. (Received February 17, 1976; )  相似文献   

11.
We discovered natural photosynthesis using Zn-containing bacteriochlorophyll in an acidophilic bacterium Acidiphilium rubrum. Chemical analysisof the cell extracts gave a 13 : 2 :1 molar ratio of Zn-bacteriochlorophyll : Mg-bacteriochlorophyll : bacteriopheophytin . Most of thepigments are associated with fully active reaction center andlight-harvesting complexes analogous to those in purple photosyntheticbacteria. The finding indicates an unexpectedly wide variabilityof photosynthesis. 7Present address: Department of Ecological Engineering, ToyohashiUniversity of Technology, Tenpaku-cho, Toyohashi, 441 Japan  相似文献   

12.
A b-type cytochrome having an -band at 560 nm was isolated fromspinach leaves (Spinacia oleracea). A method is described forpreparing this cytochrome, cytochrome b-560 (spinach), in apurified state. The cytochrome has, in its reduced state, absorption bands at560 nm (), 530 nm (ß) and 427 nm (); and in the oxidizedstate at 562 nm (), 529 nm (ß) and 417 nm (). Thepyridine ferro-haemochrome prepared from cytochrome b-560 hadan -band at 556.5 nm, indicating the protohaem-nature of theprosthetic group. The cytochrome has an oxidation-reduction potential (E'0) of+0.13V at pH 7.0, as measured using the ferri-ferro oxalate system. The cytochrome is rapidly reduced on illumination with red orfar-red light in the presence of spinach chloroplasts and isoxidized at a slower rate in the dark. This photoreduction isinhibited by 1x10–6 M 3-(3,4-dichlorophenyl)-1,1-dimethylurea(DCMU). The molecular weight of the cytochrome is 30,000 asestimated by the dextran gel filtration method. (Received December 3, 1971; )  相似文献   

13.
14.
The compositions and positional distributions of fatty acidsin the major leaf phospholipids of phosphatidylglycerol, phosphatidylcholineand phosphatidylethanolamine were analyzed by gas-liquid chromatographyand enzymic hydrolysis, and chilling-sensitive and chilling-resistantplants were comparcd with respect to the relative contents ofpalmitic and trans-3-hexadecenoic acids in the separated phospholipids.A distinct difference between these plants was found in thefatty acid compositions of phosphatidylglycerol, in which thesum of palmitic and trans-3-hexadecenoic acids ranged from 60to 78% of the total fatty acids in 8 species of chilling-sensitiveplants, and from 50 to 57% in 11 species of chilling-resistantplants. The only exception among the chilling sensitive plantsin this respect was the tomato, in which the sum of palmiticand trans-3-hexadecenic acids in phosphatidylglycerol amountedto 54%. The fatty acid compositions and the positional distributionsof fatty acids in phosphatidylglycerol suggest that the occurrenceof high proportions of dipalmitoyl and 1-palmitoyl-2-(trans-3-hexadecenoyl)species in this lipid is correlated with the susceptibilityto chilling of the leaves of higher plants. In the compositionsand positional distributions of fatty acids in phosphatidylcholineand phosphatidylethanolamine, no difference was found betweenthe chilling-sensitive and chilling-resistant plants. 1 Present address: Department of Biology, Faculty of Science,Universityof Tokyo, Hongo, Bunkyo-ku, Tokyo 113, Japan. (Received May 21, 1982; Accepted June 25, 1982)  相似文献   

15.
Tea seedlings were treated with 14C-methylamine to cause synthesisof 14C--glutamylmethylamide (N-methyl-14C). The metabolic conversionof -glutamylmethylamide was studied by tracing 14C. 14C--Glutamylmethylamide (N-methyl-14C) translocated from rootsand cotyledons to shoots of tea seedlings, was converted almostentirely into caffeine. Conversion was greater in light-exposedsamples. For those grown in the dark, the converted amount didnot correspond to the total caffeine produced. More 14C--glutamylmethylamidewas present in stems than in leaves, but with 14C-caffeine,the opposite was found. When 14C--glutamylmethylamide or 14C-methylamine was appliedto leaf disks, 14C-caffeine was biosynthesized from both substances. 1 Present address: Department of Agricultural Chemistry, ShizuokaUniversity, Iwata, Shizuoka 438, Japan. (Received September 25, 1971; )  相似文献   

16.
Uptake of the toxic heavy-metal, thallium, was studied in thecyanobacterium Synechococcus R-2 (PCC 7942) using clinicallyavailable 201Tl +. Thallium was found to distribute across theplasmalemma passively, and so the accumulation ratio of theion ([Tl+]i/[Tl+]o) could be used to calculate the apparentmembrane potential (­i,o) of the cells (ETI+i,o = ­i,o).The permeability of the plasmalemma to TI+ (PTI+ 1 to 5 nms–1)is higher than that of K+. Valinomycin does not increase thepermeability of TI+. Transient changes in the ­i,o of cells,because of electrogenic transport of ions, could be detectedfrom its effects upon the uptake rate of TI+. HCO3 hyperpolarizedSynechococcus cells, whereas NH+4, CH3NH+, and K+ led to depolarization.The use of TI+ as a reporter of ­i,o has some inherent limitations.Tl+ is toxic at very low concentrations (inhibitory effectsare apparent after about 6 h at concentrations as low as 1 mmolm–3). The rate of equilibration is slow (t1/25 to 20 min).Equilibration of TI+ takes about 2 h, which limits its valueas a membrane potential probe. Large amounts of TI+ bind tothe surface of the cells making the method impracticable formeasuring accumulation ratios of less than about 10 (­i,o)values smaller than about –60 mV). Cultures continuouslyexposed to Tl+ (10 mmol m–3) eventually become TI+ resistantby actively extruding TI+ (µTI+i,o= –3±0.2kJ mol–1) and so thallium cannot be used as a ­i,oprobe in such cells. (Received October 28, 1997; Accepted August 31, 1998)  相似文献   

17.
This work aimed to study the impacts of acquisition and assimilationof various nitrogen sources, i.e. NO3, NH4+ or NH4NO3,in combination with gaseous NH3 on plant growth and acid-basebalance in higher plants. Plants of C3 Triticum aestivum L.and C4 Zea mays L. grown with shoots in ambient air in hydroponicculture solutions with 2 mol m–3 of nitrogen source asNO3, NH4+ or NH4NO3 for 21 d and 18 d, respectively,had their shoots exposed either to 320 µg m–3 NH3or to ambient air for 7 d. Variations in plant growth (leaves,stubble and roots), and OH and H+ extrusions as wellas the relative increases in nitrogen, carbon and carboxylatewere determined. These data were computed as H+/N, H+/C, (C-A)/N,and (C-A)/C to analyse influences of different nitrogen sourceson acid-base balance in C3 Triticum aestivum and C4 Zea maysplants. Root growth in dry weight gain was significantly reduced bytreatment with 320 µg m–3 NH3 in Triticum aestivumand Zea mays growing with different N-forms, whereas leaf growthwas not significantly affected by NH3. In comparison with C3Triticum aestivum, non-fumigated C4 Zea mays had low ratiosof OH/N in NO3–3-grown plants and of H+/N in NH4+- and NH 4NO3-grown plants. Utilization of NH3 from the atmospherereduced both the OHN ratios in NO3 -grown plantsand the H+/N ratio in NH4+ - and NH4NO3 -grown plants of bothspecies. Furthermore, Zea mays had higher ratios of (C-A)/Nin NH4+ - and NH4NO3-grown plants than Triticum aestivum. Thismeans that C4 Zea mays had synthesized more organic anion perunit increase in organic N than C3 Triticum aestivum plants.Within both species, different nitrogen sources altered theratios of (C-A)/N in the order: NH4NO3>NH4+>NO3.Fumigation with NH3 increased organic acid synthesis in NO3- and NH4+ - grown plants of Triticum aestivum, whereas it decreasedorganic acid synthesis in Zea mays plants under the same conditions.Furthermore, these differences in acid-base regulation betweenC3 Triticum aestivum and C4 Zea mays plants growing with differentnitrogen sources are discussed. Key words: Acid-base balance, ammonia, ammonium, nitrate, ammonium nitrate, C3 Triticum aestivum L., C4 Zea mays L.  相似文献   

18.
Amino acid analyses of wheat -amylase purified from germinatingseeds by affinity chromatography showed a high content of amodified lysyl residue. The modified residue was identifiedas N--trimethyl lysine. The presence of trimethyl lysine in-amylase is discussed in terms of isozymes. 1 Present address: National Institutes of Health, Bldg. 10,Rm. 9B-15, Bethesda, MD 20205, U.S.A. (Received August 20, 1981; Accepted March 19, 1982)  相似文献   

19.
KUMAR  A; ELSTON  J 《Annals of botany》1992,70(1):3-9
Various kinds of measurement of tissue water status were madeseveral times during water stress and recovery in Brassica juncea(cv Canadian Black) and B napus (cv Drakkar) Unstressed plantsof the two species had similar leaf water potentials (w), solute(s) and turgor potentials (p) Values of relative water content(RWC) and the slope of the linear relationship between p andRWC (p/RWC) were greater in B napus than in B juncea Statistical correlations of pooled data for the watered andstressed treatments differentiated the relationships among RWC,w and its components in the two species The major statisticaldifference was that p/RWC was related to RWC in B napus andto w and s in B juncea A decline in p/RWC with decreasing sin B juncea may be a mechanism for maintaining p at low soilwater potentials through maintenance of more elastic cell walls. Brassica juncea, Brassica napus, osmotic adjustment, tissue elasticity, water relations  相似文献   

20.
SYNOPSIS. Studies are described on the adenosine triphosphatase(ATPase) properties of myosin isolated from skeletal muscleof Coryphaenoides, a benthic fish captured at 2,200 meters depth.Ca2+-ATPase and EDTA-ATPase of Coryphaenoides myosin show thesame pH dependence as ATPase of mammalian myosin; however, ratesof ATP hydrolysis by Coryphaenoides myosin are only 5–10%of rates of ATP hydrolysis by rabbit skeletal myosin. Coryphaenoidesmyosin ATPase shows a decrease from Q10 of 2.0 at 25°C toQ10 of 1.4 a t 2°C, and undergoes irreversible denaturationat temperatures above 25°C. At pH 6.8 to pH 8.5, Coryphaenoidesmyosin ATPase undergoes activation by pressure at 25°C,but at 2°C shows negligible effect of pressure at valuesbelow 3,000 psi. The kinetic data on Ca2+-ATPase indicate valuesof 11 kcal/mole for H, –7.5 kcal/mole for TS, and –5.7cc/mole for V at 25°C, pH 7.6. Comparable data at 2°Cindicate values of 5 kcal/mole for H. –13 kcal/mole forTS, and negligible V. According to the results of 25°C,Ca2+-activatkm of myosin-ATP may involve disruption of fouror five hydrophobic or polar groups, presumably due to an "opening-up"of the myosin molecule at or near the site for ATP binding.It would also appear that Coryphaenoides myosin has undergonean adaptive change in the enzyme mechanism for ATPase such thatthe rate of ATP hydrolysis is relatively insensitive to pressureand temperature under conditions encountered by the living fish.  相似文献   

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