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SOME PROPERTIES Of ISOLATED MITOCHONDRIAL ATPASE fROM SALMON HEART

COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY 1984年第2期79卷 263-267页

作者： THOMASSEN, JM Department of Biochemistry University of Bergen Årstadveien 19 N-5000 Bergen Norway (Tel. 291700)

A soluble Mg-dependent ATPase, similar to the mitochondrial ATPase from beef heart, is isolated from heart mitochondria of salmon (S. salar). The salmon heart ATPase has 5 subunits with MW similar to the beef heart enzyme, but the Stoke''s radius of the intact salmon enzyme is larger. The salmon heart ATPase is less temperature labile than the beef heart enzyme. The salmon heart ATPase is strongly inhibited by ADP, and the inhibition is highly temperature dependent. The ITPase activity is also inhibited by IDP (Ki = 180 .mu.m). 2,4-Dinitrophenol in small concentrations stimulates the ITPase activity and the ATPase activity of the washed salmon heart enzyme. In an enzyme preparation which had been freed of most of the bound nucleotides by dialysis in the presence of glycerol (Roveri et al., 1980) the ITPase activity is not stimulated by 2,4-dinitrophenol.

关键词： ATPase adenosine triphosphate (EC 3.6.1.4) f 1 the soluble part of the ATPase complex

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COMPARISON Of ATP-INDUCED AND DCMU-INDUCED INCREASES Of CHLOROPHYLL fLUORESCENCE

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第1期767卷 80-86页

作者： SCHREIBER, U Lehrstuhl Botanik I Universität Würzburg Mittlerer Dallenbergweg 64 D-8700 Würzburg F.R.G.

A comparative study was made (using spinach chloroplasts) of the ATP-induced and the DCMU [dichlorophenyldimethylurea] induced increases of dark chlorophyll fluorescence after activation of the latent ATPase. The ATP-induced fluorescence rise exceeds the DCMU-induced rise by an amount equivalent to the rapid component of the biphasic ATP-induced change. There is complementarity between the slower component and any preceding DCMU-induced fluorecence rise. Up to 10-4 M DCMU with the slow component being completely suppressed, the rapid ATP-induced phase is unaffected. It becomes eliminated, though, with an I50 [concentration for half-maximal inhibition] of about 3 .times. 10-4 M. No binary oscillations in dependence of the number of preilluminating flashes are observed for the rapid ATP-induced fluorescence increase. Under identical conditions such oscillations are found upon DCMU-addition. The amplitude of the rapid ATP-induced fluorescence rise is unaffected by closure of Photosystem II reaction centers in presence of DCMU and NH2OH by a single saturating flash (removal of about 50% of total quenching). With further flashes and gradual complete removal of quenching, the rapid ATP-induced change is eliminated with a 2-step dependency. The rapid phase of the ATP-induced increase in fluorescence reflects reverse electron flow at non-B-type reaction centers, while the slow phase is linked to reverse electron flow at B type centers. A model is proposed for heterogeneous interactions between the ATPase and B-type and non-B-type electron-transport chains. ''Direct coupling'' appears possible between Cf0-Cf1 [coupling factors] and those electron-transport chains which are located in the stroma-exposed margin region of the grana stacks (PS ***. units with non-B-type properties).

关键词： ATPase Chlorophyll fluorescence Photosystem II Electron transport Dichlorophenyldimethylurea (Spinach chloroplast) DCMU 3-(3′,4′-dichlorophenyl)-1,1′-dimethyl urea Q the primary acceptor of Photosystem II and quencher of chlorophyll fluorescence R or B secondary acceptor of Photosystem II PS I and PS II Photosystem I and II PQ plastoquinone f v variable fluorescence, i.e., difference between any dark-level fluorescence and maximal fluorescence P 515 field-indicating pigments of the thylakoid membrane I 50 concentration for half-maximal inhibition Cf coupling factor Cyt cytochrome

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CHARACTERIZATION Of CYTOCHROME-C-550 fROM CYANOBACTERIA

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第2期766卷 317-321页

作者： ALAM, J SPRINKLE, J HERMODSON, MA KROGMANN, DW Department of Biochemistry Purdue University West Lafayette IN 47907 U.S.A.

Cytochrome c-550 was purified from several cyanobacteria [Aphanizumenon flos-aquae, Microcystis aeruginosa, Spirulina maxima, Anabaena variabilis, Anacystis nidulans, Agmenellum quadruplicatum]. It is a low-potential, auto-oxidizable cytochrome. This cytochrome should not be confused with a degradation product of cytochrome f which may be formed during the isolation of the latter protein. Cytochromes c-550 are distinctive in size, amino-acid composition and N-terminal amino-acid sequence.

关键词： Cytochrome c -550 Cytochrome f (Cyanobacteria)

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ISOLATION, PURIfICATION AND CHARACTERIZATION Of ADENOSINE-TRIPHOSPHATASE fROM MITOCHONDRIA Of RICE SEEDLINGS

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PHYTOCHEMISTRY 1984年第12期23卷 2737-2740页

作者： BASU, A MUKHERJI, S Plant Physiology Laboratory Department of Botany University of Calcutta Calcutta 700019 India

A new and improved procedure is described for the isolation and purification of f1 ATPase from rice [Oryza sativa] seedling mitochondria. The enzyme has a multi-subunit structure as revealed by EM and SDS[sodium dodecylsulfate]-mercaptoethanol gel electrophoresis. Oligomycin sensitivity as well as cold lability of this enzyme clearly prove that the mitochondria of monocotyledons have an ATPase similar to the f1 ATPases from other souces.

关键词： Oryza sativa Gramineae rice f 1 ATPase mitochondria.

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HERBICIDE ACTION ON PHOTOSYSTEM-II IN SPINACH-CHLOROPLASTS - CONCENTRATION EffECT ON PS-II-ALPHA AND PS-II-BETA

PHOTOBIOCHEMISTRY AND PHOTOBIOPHYSICS

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PHOTOBIOCHEMISTRY AND PHOTOBIOPHYSICS 1984年第5-6期7卷 249-256页

作者： HORVATH, G DROPPA, M MELIS, A UNIV CALIF BERKELEY DIV MOLEC PLANT BIOL BERKELEY CA 94720 USA

The effect of a variety of system II herbicides on the photochemical properties of photosystem (PS) ***. and PS ***. was investigated. Urea, cyanoacrylate, phenanthroline, triazine, hydroxyquinoline, nitrophenol and benzonitrile type inhibitors were tested. At low concentrations, all herbicides were equally effective in blocking electron transport through PS ***. and PS ***.. At moderate concentrations, all herbicides selectively quenched ***. [variable fluorescence yield], the slow .beta.-component of the variable fluorescence yield emitted by PS ***.. Progressive with herbicide concentration increase, the quenching of ***., the sigmoidal and fast .alpha.-component of the variable fluorescence emission controlled by the reaction centers of PS ***. were observed. The non-variable fluorescence emission f0 remained unchanged at all herbicide concentrations. Direct measurements of the amplitude and kinetics of .delta. A320, the absorbance change of 320 nm attributed to the photoreduction of Q [primary electron acceptor of PS II], showed that, at high herbicide concentrations, the primary electron acceptor Q is unavailable for photoreduction. This phenomenon is manifested on ***. earlier than it begins affecting ***.. The differential herbicide concentration effect on fv and Q of PS ***. and PS ***. is interpreted to originate from herbicide induced disorganization of the 2 types of PS II complexes in the thylakoid membrane.

关键词： atrazine 2-chloro-4-ethylamino-6-isopropylamino-l,3,5-triazine bromoxynil 3,5-dibromo- 4-hydroxybenzonitrile DCMU 3-(3',4'-dichlorophenyl)-l, r-dimethylurea PS pholosystem Q “primary” electron acceptor of PS II f o non-variable fluorescence yield f v variable fluorescence yield,total fluorescence yield lA 320 light-induced absorbance change at 320 nm.

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ATP-INDUCED CHANGES IN THE PHOTOOXIDATION Of A LOW POTENTIAL CYTOCHROME-B-559

PHOTOBIOCHEMISTRY AND PHOTOBIOPHYSICS

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PHOTOBIOCHEMISTRY AND PHOTOBIOPHYSICS 1984年第5-6期7卷 311-317页

作者： HODGES, M PACKHAM, NK BARBER, J AFRC Photosynthesis Research Group Department of Pure and Applied Biology Imperial College London SW7 2BB U.K.

A photooxidation of the low potential form of cytochrome b-559 was observed in coupled thylakoids suspended under reducing conditions. This light-induced redox change was sensitive to inhibitors of electron flow and to uncouplers. A possible role of cytochrome b-559 LP in a Q-cycle electron transfer, in which the oxidation is driven by photosystem (PS) II and the re-reduction involves PS I is suggested. Incubation of the membranes with 500 .mu.M ATP, under conditions known to cause thylakoid protein phosphorylation, resulted in an attenuated cytochrome b-559 LP photooxidation. These changes are discussed in terms of an ATP-induced redistribution of the excitation energy between the photosystems which affects the balance of the rates of cytochrome b-559 oxidation and re-reduction.

关键词： cytochrome b-559 Q-cycle phosphorylation redox reaction absorbance change photosystem DBMIB 2,5-dibromo-3-methyl-6-isopropyl- p -benzoquinone DCMU 3,(3,4-dichloro-phenyl)-l,l-dimethylurea Cyt b -559 HP cytochrome b -559 low potential Cyt b -559 HP cytochrome b-559 high potential Cyt f cytochrome f MV methyl viologen Ant-2p 2,(3-chloro-4-trifluoro-methyl)anilino-3,5,dinitrothiophene PS photosystem

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ISOLATION AND IDENTIfICATION Of A NATURALLY-OCCURRING 7,8-DIDEMETHYL-8-HYDROXY-5-DEAZARIBOfLAVIN DERIVATIVE fROM MYCOBACTERIUM-AVIUM

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第3期797卷 377-380页

作者： NARAOKA, T MOMOI, K fUKASAWA, K GOTO, M Department of Chemistry Gakushuin University 1-5-1 Mejiro Toshima-ku Tokyo 171 Japan

A yellow pigment (C 24 H 26 N 4 PO 15 Na 3 ) was isolated from Mycobacterium avium . On the acid hydrolysis the pigment (1 mol) gave L -glutamic acid (1 mol), lactic acid (1 mol) and 7,8-didemethyl-8-hydroxy-5-deazariboflavin-5′-phosphoric acid. The structure of N-[O-[[5-(8- hydroxypyrimidol[4,5-b] quinoline -10- yl -2,4(3H, 10H)- dione )-2,3,4- trihydroxypentyloxy]hydroxyphosphoryl]- L -lactyl]- L -glutamic acid was proposed for this compound. The compound isolated functions presumably as a new cofactor in a redox system of the bacteria.

关键词： Riboflavin derivative Redox cofactor (M. avium) M-I a 7,8-didemethyl-8-hydroxy-5-deazariboflavin derivative isolated from Mycobacterium avium M-II and M-III acid hydrolytic products of M-I f 420 a redox coenzyme in methanogenic bacteria, the N-(N- l -lactyl-γ- l -glutamyl)- l -glutamic acid phosphodiester of 7,8-didemethyl-8-hyroxy-5-deazariboflavin-5′-phosphate Sf 420 7,8-didemethyl-8-hydroxy-5-deazariboflavin derivatives isolated from Streptomyces griseus

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OLIGOMYCIN SENSITIVITY-CONfERRING PROTEIN (OSCP) Of BEEf-HEART MITOCHONDRIA - INTERNAL SEQUENCE HOMOLOGY AND STRUCTURAL RELATIONSHIP WITH OTHER PROTEINS

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fEBS LETTERS 1984年第1期175卷 109-112页

作者： OVCHINNIKOV, YA MODYANOV, NN GRINKEVICH, VA ALDANOVA, NA KOSTETSKY, PV TRUBETSKAYA, OE HUNDAL, T ERNSTER, L UNIV STOCKHOLM ARRHENIUS LABDEPT BIOCHEMS-10691 STOCKHOLMSWEDEN

Structural analysis of oligomycin sensitivity-conferring protein (OSCP) revealed repeating sequences (residues 1-89, 105-190) suggesting an evolution of the protein by gene duplication. In addition to the reported homology with the delta-subunit of Escherichia coli f1ATPase, OSCP also shows a certain homology with the b-subunit of E. coli f0 and the ADP/ATP carrier of mitochondria.

关键词： f 0 f 1 Oligomycin sensitivity-conferring protein Subunit Sequence homology Structural relationship Comparative analysis

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PURIfICATION AND CHARACTERIZATION Of A NEW METALLOPROTEINASE WITH ELASTOLYTIC ACTIVITY fROM THE CATfISH PANCREAS

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第2期798卷 240-246页

作者： YOSHINAKA, R SATO, M TANAKA, H IKEDA, S KYOTO UNIV FAC AGR DEPT FISHERIES SAKYO KU KYOTO 606 JAPAN

An elastolytic enzyme was purified to homogeneity from the pancreas of the catfish P. asotus by chromatography on CM-cellulose column and affinity chromatography on (Ala)3-CH-Sepharose 4B column. The MW of the enzyme was estimated to be 24,000 by SDS[sodium dodecyl sulfate]-polyacrylamide gel electrophoresis. The enzyme had elastolytic activity was well as activity toward Suc-(Ala)3-NA [succinyl-(Ala)3-f-nitroanilide]. The enzyme was inhibited by EDTA, EGTA [ethylene glycol bis(.beta.-aminoethyl ether)-N,N,N''N''-tetraacetate], o-phenanthroline, dithiothreitol and cysteine, but not by the serine proteinase inhibitors iPR2P-f and PhCH2SO2f [diisopropylfluorophosphate and phenylmethanesulfonyl fluoride, respectively]. The enzyme was found to required Zn2+ for activity. Evidently, the catfish enzyme belongs to the group of metalloproteinases and it is a new type of pancreatic enzyme, unlike the pancreatic elastases which are serine proteinases.

关键词： Elastolytic enzyme Metalloproteinase (Catfish pancreas) iPr 2 P-f diisopropylfluorophosphate PhCH 2 SO 2 f phenylmethanesulfonyl fluoride Suc-(Ala) 3 -NA succinyl- L -alanyl- L -alanyl- L -alanine p -nitroanilide TosLysCH 2 Cl N- tosyl- L -lysylchloromethane Tos-PheCH 2 Cl N- tosyl- L -phenylalanylchloromethane EGTA ethylene glycol bis( β -aminoethyl ether)- N , N , N ′, N ′-tetraacetate Pipes 1,4-piperazinediethanesulfonic acid SDS sodium dodecyl sulfate

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CELL-VOLUME DEPENDENCE Of H-1 SPIN-ECHO NMR SIGNALS IN HUMAN-ERYTHROCYTE SUSPENSIONS - THE INfLUENCE Of INSITU fIELD GRADIENTS

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第3期803卷 137-144页

作者： ENDRE, ZH KUCHEL, PW CHAPMAN, BE UNIV SYDNEY DEPT BIOCHEM SYDNEY NSW 2006 AUSTRALIA

The 1H spin-echo NMR signal amplitudes and intensities of low MW solutes in the cytoplasm and extracellular fluid of suspensions of human erythrocytes were shown to depend on the osmotic pressure of the media. At low osmotic pressure (220 mosmol/kg), freeze-thaw lysis of the cells resulted in signal enhancement which was greatest for extracellular molecules, but both intra- and extracellular species were almost equally enhanced at 580 mosmol/kg. This effect is due to field gradients formed at cell boundaries as a result of differences in magnetic susceptibility between the medium and the cytoplasm. T2 [spin-spin relaxation time] values measured using the Carr-Purcell-Meiboom-Gill pulse sequence, with .tau. [delay time] = 0.0003 s, depended little on cell volume and absolute changes in volume magnetic susceptibility were also small. The mean field gradients, calculated from data obtained on cell suspensions at different osmotic pressures, were in the range 0.25-1.98 G[field gradient]/cm and 0.89-2.09 G/cm for intra- and extracellular compartments, respectively. The maintenance of isotonicity of the extracellular fluid during metabolic studies of cell suspensions is important in order to avoid artefacts in the determination of metabolite concentrations when using the spin-echo technique. Conversely, it may be possible to perform transport measurements using spin-echo NMR to monitor the cell volume changes which occur during the transmembrane migration of molecules.

关键词： 1 H-NMR Spin echo Cell volume field gradient (Human erythrocyte) A signal amplitude χ v volume magnetic susceptibility D translational diffusion coefficient Δδ chemical shift difference f fraction of osmotically inactive cell water f.i.d. free induction delay G ̄ mean field gradient GSH glutathione NMR nuclear magnetic resonance π osmotic pressure π 0 isotonic osmotic pressure τ delay time T 2 spin-spin relaxation time TMS tetramethylsilane V erythrocyte volume V 0 erythrocyte volume at isotonicity

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