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PARTIAL UNCOUPLING, OR INHIBITION Of ELECTRON-TRANSPORT RATE, HAVE EQUIVALENT EffECTS ON THE RELATIONSHIP BETWEEN THE RATE Of ATP SYNTHESIS AND PROTON-MOTIVE fORCE IN SUBMITOCHONDRIAL PARTICLES

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fEBS LETTERS 1985年第2期181卷 323-327页

作者： SORGATO, MC LIPPE, G SEREN, S fERGUSON, SJ CNR STUDY MITOCHONDRIAL PHYSIOL UNITI-35131 PADOVAITALY UNIV BIRMINGHAM DEPT BIOCHEMBIRMINGHAM B15 2TTW MIDLANDSENGLAND

The rates of electron transport and of ATP synthesis have been measured in bovine heart Mg-ATP submitochondrial particles oxidising succinate under conditions of partial attenuation of the proton-motive force by malonate or fCCP. This paper reports evidence that the relationship between the rate of ATP synthesis and the magnitude of the proton motive force is independent of the mode by which the decrease of the proton motive force is achieved.

关键词： ATP-synthesis rate Submitochondrial particle Proton-motive force Localised proton coupling Energy transduction Δp, proton-motive force Δμ̃ H / f = Δψ + RT 2.303/ f ·ΔpH Δψ is the electrical component and ΔpH the chemical component of the electrochemical gradient of protons (Δμ H ) existing across an energy-transducing membrane f , the faraday constant Ap 5 A, P 1 ,P 5 -di(adenosine-5'-)pentaphosphate fCCP, carbonyl cyanide p -trifluoromethoxyphenylhydrazone CCCP, carbonyl cyanide m -chlorophenylhydrazone SMP, submitochondrial particles

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A CALORIMETRY AND DEUTERIUM NMR-STUDY Of MIXED MODEL MEMBRANES Of 1-PALMITOYL-2-OLEYLPHOSPHATIDYLCHOLINE AND SATURATED PHOSPHATIDYLCHOLINES

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BIOCHIMICA ET BIOPHYSICA ACTA 1985年第2期817卷 261-270页

作者： CURATOLO, W SEARS, B NEURINGER, LJ MIT FRANCIS BITTER NATL MAGNET LAB MOLEC BIOPHYS GRP CAMBRIDGE MA 02139 USA

Binary phase diagrams were constructed from differential scanning calorimetry (DSC) data for the systems 1-palmitoyl-2-oleylphosphatidylcholine (POPC)/dimyristoylphosphatidylcholine (DMPC), POPC/dipalmitoylphosphatidylcholine (DPPC) and POPC/distearoylphosphatidylcholine (DSPC). Mixtures of POPC with DMPC exhibit complete miscibility in the gel and liquid crystalline states. Mixtures of POPC with DPPC or with DSPC exhibit gel phase immiscibility over the composition range 0-75% DPPC (or DSPC). These results, when taken together with previous studies of mixtures of phosphatidylcholines, are consistent with the hypothesis that PC whose order-disorder transition temperatures (Tm values) differ by less than ***. C exhibit gel state miscibility. Those whose Tm values differ by more than ***. C exhibit gel state immiscibility. 2H-NMR spectroscopy was used to further study mixed model membranes composed of POPC and DPPC, in which either lipid was labeled with deuterium in the 2-, 10- or 16-position of the palmitoyl chain(s) or in the N-methyls of the choline head group. POPC/DPPC mixtures in the lipid crystalline state are intermediate in order between pure POPC and DPPC at the same temperature. The POPC palmitoyl chains is always more disorderd than the palmitoyl chains of DPPC in liquid crystalline POPC/DPPC mixtures. This is attributed to the fact that a POPC palmitoyl chain is constrained by direct bonding to have at least 1 oleyl chain among its nearest neighbors, while a DPPC palmitoyl chain must have at least 1 neighboring palmitoyl chain. When liquid crystalline POPC, DPPC and POPC/DPPC mixtures are compared at a reduced temperature (relative to the acyl chain order-disorder transition), POPC/DPPC mixtures are more disordered than predicted from the behavior of the pure components, in agreement with enthalpy data derived from DSC studies. Within the temperature range of the broad phase transition of 1:1 POPC/DPPC, a superposition of gel and liquid cryst

关键词： Phospholipid membrane Mixed model membrane 2 H-NMR Differential scanning calorimetry DSC differential scanning calorimetry PC phosphatidylcholine DMPC dimyristoylPC DLPC dilauroylPC DOPC dioleylPC DPPC dipalmitoylPC DSPC distearoylPC POPC 1-palmitoyl-2-oleylPC [2- 2 H 2 ]POPC 1-[2- 2 H 2 ]palmitoyl-2-oleylPC [2,2′- 2 H 4 ]DPPC di([2- 2 H 2 ]palmitoyl)PC [10- 2 H 2 ]POPC 1-([10- 2 H 2 ]palmitoyl)-2-oleylPC [10,10′- 2 H 4 ]DPPC di([10- 2 H 2 ]palmitoyl)PC [16- 2 H 3 ]POPC 1-([16- 2 H 3 ]palmitoyl)-2-oleylPC [16,16′- 2 H 6 ]DPPC di([16- 2 H 3 ]palmitoyl)PC [ N-Me- 2 H 9 ]DPPC dipalmitoylphosphatidyl [N-Me- 2 H 9 ]choline T m order-disorder transition temperature T o transition onset temperature T f transition completion temperature

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BICARBONATE INVIVO REQUIREMENT Of PHOTOSYSTEM-II IN THE GREEN-ALGA CHLAMYDOBOTRYS-STELLATA

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JOURNAL Of PLANT PHYSIOLOGY 1985年第3期118卷 259-266页

作者： MENDE, D WIESSNER, W Abt. Expt. Phykologie Pflanzenphysiol. Inst. Univ. Göttingen Untere Karspüle 2 D-3400 Göttingen Fed. Rep. Germany.

flash induced 685 nm fluorescence emission of preilluminated and dark kept C. stellata was measured under conditions of CO2-deprivation. The difference in fluorescence intensity between dark kept and preilluminated cells is taken as a measure for the reduced state of the primary stable electron acceptor of photosystem II, Q, at the given intensity of preillumination. CO2 removal from growing cultures of this alga for 15 min diminishes photosynthetic electron transport at the oxidizing side of this photosystem. Prolonged CO2-absence influence also its reducing side. Measurements of flash induced O2 yields support the conclusion that both sides is photosystem II are affected in the absence of bicarbonate.

关键词： Chlamydobotrys stellata CO 2 -depletion fluorescence emission oxygen evolution DCMU 3(3,4-dichlorophenyl)-1,1-dimethylurea f d fluorescence emission of dark adapted algae f p fluorescence emission of preilluminated algae PQ plastoquinone PS I photosystem PS II photosystem II Q primary electron acceptor of photosystem II Y 3 oxygen flash yield of the third flash Y ss steady state oxygen evolution under flash light excitation

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MEASUREMENTS Of THE TRUE AffINITY CONSTANT IN SOLUTION Of ANTIGEN-ANTIBODY COMPLEXES BY ENZYME-LINKED IMMUNOSORBENT-ASSAY

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JOURNAL Of IMMUNOLOGICAL METHODS 1985年第2期77卷 305-319页

作者： fRIGUET, B CHAffOTTE, Af DJAVADIOHANIANCE, L GOLDBERG, ME Unité de Biochimie des Régulations Cellulaires Département de Biochimie et Génétique Moléculaire Institut Pasteur 28 rue du Docteur Roux 75724 Paris Cedex 15 France

A simple, general procedure is described for the determination of the dissociation constant (KD) of antigen-antibody equilibria in solution. first the monoclonal antibody is incubated in solution with the antigen until the equilibrium is reached; then the proportion of antibody which remains unsaturated at equilibrium is measured by a classical indirect ELISA. The experimental values of KD found by this ELISA procedure for 2 monoclonal antibodies are shown to be very close to those obtained by conventional methods (immunoprecipitation of the radiolabeled antigen, or fluorescence transfer). Moreover, it is shown that, provided the measurements are made under conditions where the total antigen concentration is in large excess over the total antibody concentration, the dissociation constant of antibody-antigen complexes can be determined even with crude preparations of monoclonal antibody. The sensitivity of the ELISA used permits the detection of very small concentrations of antibody and the determination of KD values as small as 10(-9) M. This method also offers the great advantage of dealing with unmodified molecules since no labeling of either the antigen or the antibody is required.

关键词： affinity constant monoclonal antibody ELISA tryptophan synthase β 2 β 2 -subunit of tryptophan synthase reduced holo- β 2 sodium borohydride reduced holo- β 2 -subunit f 2 proteolytic C-terminal fragment of β 2 -subunit IgG immunoglobulin G ELISA enzyme-linked immunosorbent assay EDTA ethylenediamine tetraacetic acid (di-sodium salt) IAEDANS N -iodoacetyl- N ′-(5-sulfo-1-naphtyl) ethylenediamine pyridoxal-P pyridoxal 5′-phosphate SDS sodium dodecyl sulfate PBS phosphate-buffered saline

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DIAMIDE BLOCKS H+ CONDUCTANCE IN MITOCHONDRIAL H+-ATPASE BY OXIDIZING fB DITHIOL

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fEBS LETTERS 1985年第1期192卷 83-87页

作者： HUANG, Y PRINGLE, MJ SANADI, DR HARVARD UNIV SCH MEDDEPT BIOL CHEMBOSTONMA 02115

Effects of diamide on proton conductance of electron transport particles (ETPH), purified H+-ATPase (f1-f0), f0 of the H+-ATPase from beef heart mitochondria and binding of cadmium (109Cd) to the H+-ATPase have been examined in the present paper. When ETPH and purified H+-ATPase are treated with 1 mM diamide, ATP-dependent generation of membrane potential, monitored by the absorbance change produced by the redistribution of oxonol VI, is consistently inhibited. Diamide also blocks passive H+ conductance driven by a K+ diffusion potential in the membrane sector, f0, of H+-ATPase. furthermore, diamide treatment drastically reduces the binding of 109Cd2+ to H+-ATPase, showing competition for the fB dithiol group.

关键词： Diamide Cadmium Proton conductance Sulfhydryl group f 0 H + -ATPase Mitochondria

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STIGMATELLIN - A DUAL TYPE INHIBITOR Of PHOTOSYNTHETIC ELECTRON-TRANSPORT

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BIOCHIMICA ET BIOPHYSICA ACTA 1985年第2期807卷 216-219页

作者： OETTMEIER, W GODDE, D KUNZE, B HOfLE, G GESELL BIOTECHNOL FORSCH MBH MIKROBIOL ABTD-3300 BRUNSWICKFED REP GER GESELL BIOTECHNOL FORSCH MBH NAT STOFFCHEM ABTD-3300 BRUNSWICKFED REP GER

Stigmatellin, an antibiotic produced by the myxobacterium Stigmatella aurantiaca , is a powerful inhibitor of photosynthetic electron transport. In the photosynthetic electron-transport chain, it has two different inhibition sites. One is located at the reducing side of Photosystem II ( I 50 value, 52.5 nM) and identical to the inhibition site of inhibitors of the 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea type. The second one is located at the cytochrome b 6 f- complex ( I 50 value, 59.0 nM) and corresponds to the inhibition site of 2.5-dibromo-3-methyl-6-isopropyl-1,4-benzoquinone. Stigmatellin is the most potent inhibitor of spinach cytochrome b 6 f complex known so far.

关键词： Photosystem II Cytochrome b 6 f complex Inhibitor Stigmatellin (Spinach chloroplast) azido-atrazine 2-azido-4-(ethylamino)-6-(isopropylamino)- s -triazine Chl chlorophyll DBMIB 2,5-dibromo-3-methyl-6-isopropyl-1,4-benzoquinone DCIP 2,6-dichlorophenolindophenol DCMU 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea DNP-INT 2-iodo-2′,4,4′-trinitro-3-methyl-6-isopropyl-diphenylether Hepes 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid Mes 2-( N -morpholino)ethanesulfonic acid metribuzin 4-amino-6- t -butyl-3-(methylthio)-1,2,4-triazin-5-(4H)one UHDBT 5- n -undecyl-6-hydroxy-4,7-dioxobenzothiazole

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EffECTS Of CERULENIN ON THE PHOTOSYNTHETIC PROPERTIES Of DETACHED BARLEY LEAVES DURING CHLOROPLAST DEVELOPMENT

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JOURNAL Of PLANT PHYSIOLOGY 1985年第1期119卷 55-64页

作者： LASKAY, G LEHOCZKI, E SZALAY, L Department of Biophysics József Attila University Szeged Hungary

The effects of cerulenin on the accumulation of photosynthetic pigments and on the development of photosynthetic activity during the greening of excised etiolated barley leaves were studied. Cerulenin at 1.5 .times. 10-4 M caused 70% inhibition of the accumulation of chlorophyll b, 50% of that of chlorophyll a, and 40% of that of total carotenoids after 72 h of greening. Cerulenin treatment also prevented the further increase of electron transport activity from H2O to K-ferricyanide after 24 h of greening, whereas the activity of photosystem-I was unaffected by cerulenin during greening. The ability of ammonium chloride to uncouple electron transport and photophosphorylation increased with the duration of greening in untreated chloroplasts. This tendency was also distributed in the chloroplasts from cerulenin treated leaves. The greening process was slowed down in cerulenin-treated leaves, as revealed by fluorescence spectra and fluorescence induction measurements. The rate of electron transport from the primary electron acceptor of photosystem-II to the secondary acceptor pool was also decreased in intact cerulenin-treated leaves, this causing a more than 30% inhibition of the steady state CO2 fixation on a chlorophyll basis of intact leaves after 72 h of greening in the presence of cerulenin.

关键词： Hordeum vulgare cerulenin chloroplast development electron transport fluorescence induction ACP acyl carrier protein DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea DCPIP 2,6-dichlorophenol-indophenol f o constant or «dead» fluorescence f v variable fluorescence f rel relative fluorescence intensity PS-I photosystem-I PS-II photosystem-II Q «primary» electron acceptor of photosystem-II (also denoted to «quencher»)

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LEUKEMIA X fIBROBLAST HYBRID-CELLS PROLONG THE LIVES Of LEUKEMIC MICE

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EUROPEAN JOURNAL Of CANCER & CLINICAL ONCOLOGY 1985年第5期21卷 637-643页

作者： WANG, DR SLOMSKI, R COHEN, EP SICHUAN MED COLL DEPT MICROBIOL CHENGDU PEOPLES R CHINA POLISH ACAD SCI INST HUMAN GENET PL-60479 POZNAN POLAND UNIV ILLINOIS DEPT MICROBIOL & IMMUNOL CHICAGO IL 60680 USA

ASL-1 leukemia .times. LM(TK-) fibroblast hybrid cells prolong the lives of leukemic (A/J .times. C3H/HeJ)f1 mice. The hybrid cells, like the fibroblast cells used in forming the hybrid, have lost malignant growth properties in immunocompetent recipients and are rejected. Mice receiving hybrid cells along with ASL-1 cells exhibit immunity toward the leukemia cells;approximately 50% of the animals injected with 106 or more hybrid cells along with ASL-1 cells survive > 60 days;animals in the control group injected with leukemia cells alone invariably die in shorter intervals. The immunity generated is persistent for at least 6 mo. Some leukemic mice receiving doses of combination chemotherapy which are insufficinet to cure them of the disease survive for prolonged and at times indefinite periods if they are injected with hybrid cells. The immunity generated in mice receiving hybrid cells is directed toward a leukemia-associated antigen of leukemia cells expressed by hybrid cells as well. In mixed lymhphocyte culture a heightened stimulation of spleen cells from hybrid cell-injected mice toward ASL-1 cells is observed.

关键词： Hybrid cells ASL-1×LM(TK-) cells f 1 mice (A/J×C3H/HeJ)f 1 mice i.p. intraperitoneal MLR mixed lymphocyte reaction cytosine arabinoside and methotrexate combination chemotherapy s.c. subcutaneously

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RELATIONSHIP Of AUER RODS AND CHROMOSOME fINDINGS TO OUTCOME IN 89 ADULTS WITH ACUTE NONLYMPHOBLASTIC LEUKEMIA

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LEUKEMIA RESEARCH 1985年第10期9卷 1231-1235页

作者： KRIS, MG MERTELSMANN, R JHANWAR, S CHAGANTI, R SZATROWSKI, TH GEE, TS ARLIN, Z KEMPIN, S BENEDETTO, P CLARKSON, B CORNELL UNIV MED CTRCOLL MEDMEM SLOAN KETTERING CANC CTR DEPT MEDHEMATOL LYMPHOMA SERV NEW YORK NY 10021 USA CORNELL UNIV MED CTRCOLL MEDMEM SLOAN KETTERING CANC CTR DEPT PATHOL NEW YORK NY 10021 USA RUTGERS STATE UNIV GRAD SCH MANAGEMENT NEWARK NJ 07102 USA

We performed cytogenetic analyses using banding techniques on 89 adults with acute nonlymphoblastic leukemia prior to receiving protocol chemotherapy. The relationships of cytogenetic findings both to outcome and to other pretreatment variables (particularly the presence or absence of Auer rods) were analyzed. Patients were followed up to 90 + months. When patients were grouped according to cytogenetic findings (NN: all normal metaphases;AA;all abnormal metaphases;AN: both normal and abnormal metaphases;f: no evaluable metaphases;I: insufficient (less than three) metaphases) no significant differences were noted with regard to sage, sex, terminal transferase positivity, complete remission rate, remission duration or survival. The marrow aspirates of patients with only normal (69%) metaphases or no evaluable metaphases (64%) were more likely to display Auer rods than specimens for individuals with only abnormal (26%) or a mixture of normal and abnormal (42%) metaphases (p = 0.03). The presence of Auer rods in the pretreatment marrow aspirate was associated with an increased complete remission rate (71% vs 41%, p = 0.004), median remission duration (12 months vs 9 months, p = 0.02), and median survival (13 months vs 4 months, p = 0.01). Using multivariable analyses, the presence or absence of Auer rods was the pretreatment factor that most significantly predicted response and survival in this group of patients. The presence of a normal karyotype in the initial cytogenetic preparation is associated with the presence of Auer rods. The findings of Auer rods in the initial bone marrow predicts greater response and longer survival in acute nonlymphoblastic leukemia.

关键词： Acute nonlymphoblastic leukemia cytogenetics Auer rods chromosomes prognostic factors ANLL acute nonlymphoblastic leukemia CR complete remission AA abnormal metaphases only AN abnormal and normal metaphases I insufficient numbers of metaphases f failure of cell growth — no metaphases NN normal metaphases only fAB french-American-British

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INTERACTION BETWEEN THE MITOCHONDRIAL ATP-SYNTHETASE AND ATPASE INHIBITOR PROTEIN - ACTIVE INACTIVE SLOW PH-DEPENDENT TRANSITIONS Of THE INHIBITOR PROTEIN

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fEBS LETTERS 1985年第2期184卷 226-230页

作者： PANCHENKO, MV VINOGRADOV, AD MOSCOW STATE UNIV SCH BIOLDEPT BIOCHEMMOSCOW 119899USSR

The rate of mitochondrial ATPase inactivation by the naturally occurring inhibitor protein in the presence of saturating ATP and Mg2+ at pH 8.0 depends hyperbolically on the amount of inhibitor added; the upper limit of an apparent first-order constant for the inactivation process is 1.0(-1) at 25 degrees C. A dramatic difference in the inactivation rate is observed when the protein inhibitor is added to the same assay system from either acidic (pH 4.8) or alkaline (pH 8.2) solutions. The slow reversible transition of the inhibitor from its rapidly reacting 'acidic' form to the slow reacting 'alkaline' form occurs when the solution of the protein inhibitor is subjected to a pH-jump from 4.8 to 8.2 (t1/2 approximately 30s at 25 degrees C). The pH-profile of the inhibitor active/inactive equilibrium suggests that a group with pKa approximately 6.5 is involved in the transition. Treatment of the inhibitor protein with a histidine-specific reagent (e.g. diethyl pyrocarbonate) abolishes its inactivating effect on the ATPase activity. It is concluded that the protonation/deprotonation of the inhibitor protein followed by its slow conformational changes is the rate-limiting step in the inhibitor-ATP synthetase interaction.

关键词： ATP synthetase Inhibitor protein Acid-base transition Mitochondria f 1 , beef heart mitochondrial coupling factor If 1 , beef heart ATPase protein inhibitor AS-particles, submitochondrial ultrasonic particles depleted of If 1 by slow filtration through Sephadex Mops, 3-( N -morpholino)propanesulphonate Mes, 4-morpholineethanesulphonate fCCP, carbonyl cyanide p -trifluoromethoxyphenylhydrazone

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