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ISOLATION AND PARTIAL CHARACTERIZATION Of A ALPHA-2-BETA-1-GLYCOPROTEIN fROM HORSE PLASMA

INTERNATIONAL JOURNAL Of BIOCHEMISTRY 1983年第8期15卷 1013-1018页

作者： PELLEGRINI, A ZWEIfEL, HR VONfELLENBERG, R Department of Veterinary Physiology University of Zurich Winterthurerstr 260 8057 Zürich Switzerland

A .alpha.2-.beta.1-glycoprotein was isolated from horse plasma by classical methods. The final product appeared homogeneous by agarose gel and pore limit SDS[sodium dodecyl sulfate] polyacrylamide gel electrophoresis, immunoelectrophoresis and crossed immunoelectrophoresis. The protein moved in agarose gel electrophoresis just above the .beta.1 region and seemed composed of a single polypeptide chain. A highly heterogenic banding pattern, focused between pH 5.1 and 6.5 was revealed by isoelectric focusing. The MW determined by gel filtration on Sephadex G100 and by a pore limit polyacrylamide gel electrophoresis in presence of SDS were 65,000 and 82,300 dalton, respectively. No serological relation was found between the horse .alpha.2-.beta.1-glycoprotein and human and bovine plasma proteins.

关键词： SDS-sodium dodecyl sulphate BIS—methylenebisacrylamide TEMED- N , N ′, N ′, N ′-tetramethy-lenediamine K 2 EDTA-ethylenediaminetetraacetic acid dipotassium salt Affi Gel Blue-Cibacron Blue f3GA crossed-linked to Bio Gel® A5-M 2-ME-2 mercaptoethanol f α 1 -PI-fast α 1 -protease inhibitor T -Total acrylamide concentration C -methylenebisacrylamide concentration

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ENERGETICS Of PHOTOSYNTHESIS IN ZEA-MAYS .2. STUDIES Of THE fLASH-INDUCED ELECTROCHROMIC SHIfT AND fLUORESCENCE INDUCTION IN MESOPHYLL CHLOROPLASTS

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BIOCHIMICA ET BIOPHYSICA ACTA 1983年第1期722卷 127-136页

作者： CROWTHER, D LEEGOOD, RC WALKER, DA HIND, G UNIV SHEFFIELD DEPT BOT ARC PHOTOSYTHN RES GRP SHEFFIELD S10 2TN S YORKSHIRE ENGLAND BROOKHAVEN NATL LAB DEPT BIOL UPTON NY 11973 USA

Patterns of electron transfer in isolated mesophyll chloroplasts of maize (Z. mays L.) were studied in the presence of the physiological substrates, oxaloacetate, 3-phosphoglycerate and pyruvate. flash-induced absorbance changes due to the electrochromic pigment band-shift (P-518) were used to estimate relative electron flow rates through the cyclic and noncyclic pathways of electron transport. further information on the redox state of electron carriers and the activity of coupled electron flow was obtained from measurements of fluorescence induction and of actinic-light-induced fluorescence changes. The importance of correct redox poising for optimal rates of photosynthesis was demonstrated and the operation and regulation of photosynthesis in the C4 system discussed.

关键词： Photosynthesis Electron transport Chlorophyll fluorescence fluorescence induction (Zea mays mesophyll chloroplast) DCMU 3-(3,4-dichlorophenyl)-1, 1-dimethylurea Mes 2-( N -morpholino)ethanesulphonic acid Tricine N -tris(hydroxymethyl)methylglycine f 0 intrinsic bed fluorescence f i initial level of fluorescence seen during rapid induction ( f 0 plus a variable component proportional to the state of reduction of Q) f m maximal level of fluorescence (seen in the presence of DCMU) f v variable fluorescence (maximally f m − f 0 ) Chl chlorophyll PS photosystem

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THE ASSOCIATIONS Of MEMBRANE SYSTEMS

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BIOELECTROCHEMISTRY AND BIOENERGETICS 1983年第4期10卷 315-333页

作者： BOURD, GI MARTIROSOV, SM ALL UNION GENET RES INST BOX 45EREVAN 375056ARSSR NF GAMALEYA EPIDEMIOL & MICROBIOL INST MOSCOW 123098USSR

This review is devoted to the data on the intramembranal interactions of proteins for the regulation of membrane transport, of cell metabolism anf of energy transformation. first discussed are the results on H + transfer via the ATPase complex f 1 f 0 and the counter-transport of K + in glycolysing bacteria. It is concluded that the f 1 f 0 and the natural K + ionophore are able to form an intramembranal binary supercomplex for the joint employment of ATP energy. Such a supercomplex operates like an ionic pump, exchanging 2 H + for K + and capable of both hydrolysis and synthesis of ATP with the essential decrease of energy dissipation. It is also assumed that such a system in gram-negative bacteria includes, as a third portion, a protein regulator which turns the H + /K + exchange on and off dependent on osmotic pressure in the medium. The data on H + /K + antiport in bacteria are also considered. In this case the possibility of the formation of another type of supercomplex formed by the proton channel f 0 and the K + ionophore is suggested. Thus, the interaction of membrane proteins for the joint employment of converted energy ATP, Δ μ H + and Δ μ K + is considered. In the second part of the review the results on the cell metabolism regulation through the intramembranal interaction of some systems in bacteria are described. These regulations are performed via the mechanisms known as “ inducer exclusion ” and “ catabolite repression ”. The reciprocal inhibiting effect under the interplay of PTS and lactose permease is shown. The lactose permease is assumed to be inoperative due to the interaction with E 11 when the latter translocates glucose (or mannose). Some experiments serve as the evidence that the catabolite represssion connected with the fall in adenylate cyclase activity occurs at membrane level. The transport of glucose through the enzyme E 11 or lactose via M-protein leads to the inhibition of the cyclase function as a result of intramembranal interaction

关键词： DCCD N,N′-dicyclohexylcarbodiimide Δ μ is the electrochemical potential difference Δψ is the membrane potential f 1 f 0 is the ATPase complex for H + transport f 1 is the ATPase f 0 the proteolipid incorporated in a membrane TrkA is the system for transfer of K + Trkf is the H + /K + -exchanging system ΔpH and Δp K are H + and K + gradients respectively PTS is the phosphoenolpyruvate-dependent carbohydrate phosphotransferase system PEP is phosphoenolpyruvate MG is methyl-α- d -glucopyranoside AC is adenylate cyclase M-protein is β-galactoside permease, product of gene lac Y ONPG is o -nitrophenyl-β- d -galactopyranoside NEM is N-ethylmaleimide TDG is thiodigalactoside E 11 is the membrane component of the PTS.

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THE EffECT Of DDT AND RELATED INSECTICIDES ON THE MITOCHONDRIAL ATPASE Of HOUSEfLIES

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COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY 1983年第2期74卷 259-266页

作者： CHEfURKA, W Agricultural Research Centre Agriculture Canada University Sub P.O. London Ontario Canada N6A 5B7

DDT is a weak inhibitor of the ATPase of housefly muscle mitochondria in the absence of Mg2+ but an activator in the presence of Mg2+. By contrast, DDT and several p,p''-substituted .alpha.-trichlomethylbenzylanilines were strong inhibitors of the ATPase activity in the presence of the uncoupler, dinitrophenol. Maximum inhibition of the DNP-ATPase was achieved when the ATPase complex was dissociated from its endogenous protein inhibitor. The inhibition by DDT was noncompetitive, maximum at acid pH and independent of temperature. The inhibition was counteracted by exogenous phosphatidylcholine and phosphatidylethanolamine. The ATPase was also activated by NH4+ in the presence of valinomycin. This activation was reversed by K+ and strongly inhibited by DDT. The possible mechanisms involved in the inhibition by DDT are discussed.

关键词： DDT 2,2- bis ( p -chlorophenyl)1,1,1-trichloroethane ATP adenosine triphosphate DNP 2,4-clinitrophenol EDTA ethylenediamine tetraacetic acid EGTA ethyleneglycol-bis(B-aminoethyl) ether N , N '-tetraacetic acid Tris tris(hydroxymethyl)aminoethane f 1 soluble mitochondrial ATPase f 0 factor conferring oligomycin sensitivity

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SURVEY Of PLANT INHIBITORS Of POLYMORPHONUCLEAR LEUKOCYTE ELASTASE, PANCREATIC ELASTASE, CATHEPSIN-G, CATHEPSIN-B, HAGEMAN-fACTOR fRAGMENTS, AND OTHER SERINE PROTEINASES

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BIOCHEMICAL PHARMACOLOGY 1983年第6期32卷 985-990页

作者： HOJIMA, Y PISANO, JJ COCHRANE, CG SCRIPPS CLIN & RES FDN DEPT IMMUNOPATHOL LA JOLLA CA 92037 USA NHLBI CHEM LAB BETHESDA MD 20205 USA

Various flower bulbs and vegetable and legume seeds were tested for inhibitors of polymorphonuclear leukocyte elastase, pancreatic elastase, cathepsin G, cathepsin B, trypsin, .alpha.-chymotrypsin, Hageman factor fragments, plasma kallikrein and plasmin. Calla bulbs contained a 33,000 dalton polymorphonuclear leukocyte elastase inhibitor and a 4,000 dalton cathepsin G inhibitor. Seeds of some members in the Cruciferae family, such as radish and broccoli, contained one or more 2500-4000 dalton inhibitors which inhibited cathepsin G, trypsin, Hageman factor fragments and plasmin, but not plasma kallikrein. These seeds also contained a 1000 dalton cathepsin B inhibitor. The above inhibitors were probably polypeptides which inhibited proteinases by making an enzyme-inhibitor complex, with the exception of the cathepsin B inhibitor. These newly found inhibitors with their characteristic profiles of inhibition should be useful in biochemical and pathophysiological studies on granulocyte proteinases and enzymes of the coagulation and fibrinolytic pathways.

关键词： PMN polymorphonuclear leukocyte MeO methoxy Suc succinyl p NA p -nitroanilide Bz, benzoyl BSA bovine serum albumin Hf f Hageman factor fragments STI soybean trypsin inhibitor LTI lima bean trypsin inhibitor PTI pancreatic trypsin inhibitor IU inhibitor unit

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CHARACTERIZATION Of CHLOROPHYLL fLUORESCENCE QUENCHING IN CHLOROPLASTS BY fLUORESCENCE SPECTROSCOPY AT 77-K .1. DELTA-PH-DEPENDENT QUENCHING

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BIOCHIMICA ET BIOPHYSICA ACTA 1983年第2期723卷 169-175页

作者： KRAUSE, GH BRIANTAIS, JM VERNOTTE, C CNRS PHOTOSYNTHESE LAB F-91190 GIF SUR YVETTE FRANCE

The nature of the light-induced .DELTA. pH-dependent decline of chlorophyll a fluorescence (f) in intact and broken spinach chloroplasts was investigated. fluorescence spectra at ***. K of chloroplasts frozen in the low-fluorescent (high .***) state showed increased ratios of the bank peak at 735 nm (Photosystem (PS) I fluorescence) to the peak at 695 nm (PS II fluorescence). The increase in the f735/f695 ratio at ***. K was related to the extent of fluorescence quenching at room temperature. Normalization of low-temperature spectra with fluorescein as an internal standard revealed a lowering of f695 that was not accompanied by an increase in f735: preillumination before freezing decreased both f695 and, to a lesser extent, f735 in the spectra recorded at ***. K. fluorescence induction of chloroplasts frozen in the low-fluorescent state showed a markedly decreased variable fluorescence (fv) of PS II, but no concomitant increase in initial fluorescence (f0) of PS I. Thus, the buildup of a proton gradient at the thylakoid membrane, as reflected by fluorescence quenching at room temperature, affects low-temperature fluorescence emission in a manner entirely different from the effect of removal of Mg2+, which is thought to alter the distribution of excitation energy in favor of PS I. The .***-dependent quenching therefore cannot be caused by such change in energy distribution and is suggested to reflect increased thermal deactivation.

关键词： Chlorophyll fluorescence fluorescence quenching Proton gradient (Spinach chloroplast) Chl chlorophyll PS photosystem f fluorescence emission P peak S steady state of fluorescence emission Q primary electron acceptor of PS II Hepes N -2-hydroxyethylpiperazine- N ′-2-ethanesulfonic acid

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STOICHIOMETRY Of THE H+-ATPASE Of ESCHERICHIA-COLI-CELLS DURING ANAEROBIC GROWTH

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fEBS LETTERS 1983年第2期154卷 343-346页

作者： KASHKET, ER Department of Microbiology Boston University School of Medicine Boston MA 02118 USA

The H+/ATP stoichiometry of the H+-ATPase was investigated in Escherichia coli cells growing under anaerobic conditions at pH 6 and 7. The protonmotive force was determined from the intracellular accumulation of benzoate and tetraphenylphosphonium ions, as well as the accumulation of lactose in this lac operon inducible, but beta-galactosidase negative strain. The phosphorylation potential was calculated from the cellular concentrations of ATP, ADP and inorganic phosphate. By comparing the phosphorylation potential and the proton motive force under these steady state conditions, the H+/ATP stoichiometry was determined to be 3, similar to the value previously found in the same cells growing under aerobic conditions.

关键词： H + -ATPase Proton motive force Escherichia coli Phosphorylation potential Anaerobiosis Stoichiometry EDTA ethylenediaminetetraacetic acid TPP + tetraphenylphosphonium ion Tris tris(hydroxymethyl)aminomethane XDP xanthosine diphosphate PMf or Δp protonmotive force = Δψ − Z ΔpH = Δ≈μ H + / f Δψ transmembrane electrical gradient ΔpH transmembrane chemical gradient of H + = pH out (pH of the bulk medium)-pH in (pH of the cytosol) Δp lac Δμ lac /f ΔG′ p phosphorylation potential = Gibbs free energy of ATP synthesis = ΔG′ 0 + RT ln[ATP]/[ADP][P i ] ΔG′ 0 standard free energy of ATP hydrolysis

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INTERACTION Of POLYMERASES WITH 2'-DEOXYURIDINE-5'-TRIPHOSPHATE SPIN-LABELED AT THE 5-POSITION

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fEBS LETTERS 1983年第2期152卷 157-162页

作者： WARWICKKOOCHAKI, PE HAKAN, A BOBST, AM Department of Chemistry University of Cincinnati Cincinnati OH 45221 USA

2'-Deoxyuridine-5'-triphosphate spin-labeled at the 5-position with N-[1-oxyl-2,2,6,6-tetramethyl-4-piperidinyl]-O- was found to be an inhibitor of some DNA and RNA polymerases including avian myeloblastosis virus reverse transcriptase. furthermore, the spin-labeled nucleotide was found to be incorporated internally into polydeoxythymidylic acid via reverse transcriptase to an extent of 1.0 spin-labeled base per 10(3) bases. The incorporation, monitored by electron spin resonance, is analogous to some other nucleotide inhibitors of polymerases, and the results indicate that it may be feasible to obtain sequence specific, spin-labeled DNA, enzymatically.

关键词： Spin-labeled nucleotide Reverse transcriptase inhibition DNA polymerase f 3 dTTP 5-trifluoromethyl-2′-deoxyuridine-5′-triphosphate araCTP cytosine-D-arabino-furanoside-5′-triphosphate pppDUGT 2′-deoxy-uridine-5′-triphosphate substituted at the 5-position with N -[1-oxyl-2,2,6,6-tetramethyl-4-piperidinyl]-0- AMV avian myeloblastosis virus ESR electron spin resonance

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STIMULATION Of A CYCLIC ELECTRON-TRANSfER PATHWAY AROUND PHOTOSYSTEM-II BY PHOSPHORYLATION Of CHLOROPLAST THYLAKOID PROTEINS

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fEBS LETTERS 1983年第1期162卷 81-84页

作者： HORTON, P LEE, P UNIV SHEFFIELD ARCPHOTOSYNTHESIS RES GRPSHEFFIELD S10 2TNS YORKSHIREENGLAND

fluorescence induction kinetics of phosphorylated and unphosphorylated chloroplast thylakoid membranes have been studied under low light intensity. Hydroxylamine is able to enhance the fluorescence yield to a greater extent in phosphorylated membranes in both the presence and absence of DCMU. Examination of the light intensity dependence of the fluorescence yields leads to the conclusion that protein phosphorylation enhances the efficiency of a NH 2 OH-sensitive cyclic electron-transfer pathway around photosystem II.

关键词： Photosynthesis Chloroplast Protein phosphorylation Thylakoid Photosystem Chlorophyll fluorescence PSII photosystem II PSI photosystem I Q the primary acceptor of PSII whose redox state controls the yield of fluorescence f o the level of fluorescence when Q is oxidised f mv f m - f o , the variable fluorescence DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea LHC-II the light harvesting complex associated with PSII

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DIOXYGEN ACTIVATION IN TRANSITION-METAL COMPLEXES IN THE LIGHT Of MOLECULAR-ORBITAL CALCULATIONS

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COORDINATION CHEMISTRY REVIEWS 1983年第1-2期50卷 1-72页

作者： BOCA, R Department of Inorganic Chemistry Slovak Technical University CS-812 37 Bratislava Czechoslovakia

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