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fREEZE-fRACTURE STUDIES ON BARLEY PLASTID MEMBRANES .8. IN VIRIDIS-115, A MUTANT COMPLETELY LACKING PHOTOSYSTEM-II, OXYGEN EVOLUTION ENHANCER-1 (OEE1) AND THE ALPHA-SUBUNIT Of CYTOCHROME-B-559 ACCUMULATE IN APPRESSED THYLAKOIDS

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BIOCHIMICA ET BIOPHYSICA ACTA 1989年第1期975卷 164-174页

作者： SIMPSON, DJ VALLON, O VONWETTSTEIN, D Department of Physiology Carlsberg Laboratory Copenhagen Valby (Denmark)

The barley nuclear gene mutant viridis-115 completely and specifically lacks Photosystem II (PS II)-dependent electron-transport activity. This was associated with the absence of variable fluorescence in room-temperature fluorescence induction, and a change in the 77 K fluorescence emission spectrum due to fluorescence from light-harvesting chlorophyll a/b-protein of Photosystem II (LHC II) unquenched by PS II reaction centers. The low-temperature absorption spectrum lacked chlorophyll a absorbing at 683 nm. Thin section electron microscopy showed that the grana were about 5 times larger in diameter than the wild type, with almost no inter-grana lamellae. Quantitation of the freeze-fracture ultrastructure of viridis-115 revealed a 96% reduction in the number of endoplasmic fracture face of thylakoid (Efs) particles, the presumed site of the PS II reaction centers. No change was seen in the protoplasmic fracture face of stacked (Pfs) or unstacked (Pfu) thylakoid particle densities, and the mutant contained 68% of the wild-type density of endoplasmic fracture face of unstacked thylakoid (Efu) particles, making it unlikely that they represent PS II located in the stroma lamellae (PS ***.). Calculations based on chlorophyll:P-700 and chlorophyll a/b ratios revealed a loss of between 60 and 70 molecules of chlorophyll a per PS II reaction center. A double mutant, clo-f22800 .times. vir115 contained grana in the absence of PS II and LHC II, both of which have been implicated in the maintenance of thylakoid adhesion. Three of the ''core'' PS II polypeptides (CP47, CP43 and D1) were severely deficient, whereas the .alpha.-subunit of cytochrome b-559 was present in near normal levels, although the high potential form was not detectable by spectroscopy. The chlorophyll a/b-proteins CP-29 and LHC1 were present at normal levels, as was the extrinsic 33 kDa oxygen evolution enhancer 1 (OEE1).

关键词： Absorption spectroscopy Chlorophyll protein Chloroplast Electron microscopy fluorescence emission Immunocytochemistry P-700 CP chlorophyll-protein DPC 1,5-diphenylcarbazide DCIP 2,6-dichlorophenylindophenol Efs/u endoplasmic fracture face of stacked/unstacked thylakoids ESs endoplasmic surface of stacked thylakoids f 0 fluorescence yield at onset of illumination f m maximum yieldof fluorescence f v variable fluorescence HP high potential LP low potential LHC I (II) light-harvesting chlorophyll a/b -protein of Photosystem I (II) LP low potential OEE oxygen evolution enhancer PAGE polyacrylamide gel electrophoresis Pfs/u protoplasmic fracture face of stacked/unstacked thylakoids PS I (II) Photosystem I (II) SDS sodium dodecyl sulphate

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COMPARISON Of PROPRANOLOL-BINDING PLASMA-PROTEINS IN SHEEP WITH THOSE IN HUMANS, DOGS AND RATS

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BIOCHEMICAL PHARMACOLOGY 1989年第23期38卷 4199-4205页

作者： HILL, MD BRISCOE, PR ABRAMSON, fP GEORGE WASHINGTON UNIV MED CTR DEPT PHARMACOL 2300 EYE ST NW WASHINGTON DC 20037 USA

Alpha1-acid glycoprotein (AGP), a plasma protein responsible for the binding of a variety of basic lipophilic drugs including propranolol, is different from other plasma proteins in being nonprecipitable after treatment with 1.2 M perchloric acid (PCA). To assess the contribution of AGP to drug disposition in sheep and three other species (rats, dogs, and humans), the binding of [3H]propranolol was measured before and after PCA precipitation. PCA precipitation reduced propranolol binding 14-fold in sheep, compared to 2- to 3-fold in the other species. This implied either that sheep AGP binds less propranolol than other species, or that the AGP in sheep is more precipitable. It was not due to inherently poor propranolol binding, as whole sheep plasma bound a higher fraction than the other species. When samples of PCA-precipitation sheep plasma were anlayzed using polyacrylamide gel electrophoresis, the concentration of AGP was 10-20% that of the other species. Phenobarbital induction was used as a tool to examine the changes in the plasma protein profile. Phenobarbital induced propranolol binding and AGP along with two other proteins in sheep. One of these proteins migrated similarly to AGP deglycosylated by peptide-N-glycosidase f. It is postulated that the greater precipitability of propranolol binding in sheep is due to a less glycosylated form of AGP which is not important in other species.

关键词： AGP α 1 -acid glycoprotein PCA perchloric acid B f ratio of bound to free concentrations of the drug PAGE polyacrylamide gel electrophoresis SDS sodium dodecyl sulfate PNGase peptide- N -glycosidase f

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SPECTROSCOPIC INVESTIGATIONS Of fLUORESCENCE BEHAVIOR, ROLE AND fUNCTION Of THE LONG-WAVELENGTH PIGMENTS Of PHOTOSYSTEM-I

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BIOCHIMICA ET BIOPHYSICA ACTA 1989年第2期973卷 198-204页

作者： STAHL, U TUSOV, VB PASCHENKO, V VOIGT, J HUMBOLDT UNIV DEPT PHYS DIV EXPTL BIOPHYS & SPECTROSCOPY INVALIDENSTR 42 DDR-1040 BERLIN GER DEM REP MV LOMONOSOV STATE UNIV DEPT BIOL MOSCOW 117234 USSR

A method was established for quantitative evaluation of fluorescence spectra of intact leaves without expensive sample preparation. fluorescence induction and excitation effects were used to evaluate the contribution of Photosystem I to the long-wavelength fluorescence of intact leaves at room temperature which appeared to be about 25%. To characterize its spectral line-shape we studied low-temperature fluorescence. The connection of these three methods made it possible to determine the quantitative spectra of both photosystems in intact leaves. Investigations of fluorescence decay kinetics on isolated Photosystem I subchloroplast particles provided information about functional aspects of the long-wavelength pericentral antenna pigments. It appeared that these pericentral pigments are not essential for transferring excitation energy toward photochemically active reaction centres. Only if the reaction centres were closed the pericentral antenna would act as a protection guard against photodestruction.

关键词： Chlorophyll fluorescence Photosystem I Chi chlorophyll f xyz fluorescence at xyz nm LHA light-harvesting antenna PS Photosystem RC reaction centre λ E excitation wavelength

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IMPORTANCE Of MULTIPLE HYDROXYLATED METABOLITES IN HEXAMETHYLPHOSPHORAMIDE (HMPA)-MEDIATED MUTAGENESIS IN DROSOPHILA-MELANOGASTER

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MUTATION RESEARCH 1989年第2期212卷 193-211页

作者： ZIJLSTRA, JA BRUSSEE, J VANDERGEN, A VOGEL, EW STATE UNIV LEIDEN DEPT RADIAT & CHEM MUTAGENESIS 2333 AL LEIDEN NETHERLANDS STATE UNIV LEIDEN DEPT CHEM 2333 AL LEIDEN NETHERLANDS

The mutagenic profiles in Drosophila and the influence of inhibition of metabolism on genotoxic activity were determined for hexamethylphosphoric triamide (HMPA), some synthetically prepared presumed metabolites and ethylated analogs. Demethylated HMPA metabolites are considerably less mutagenic than HMPA, dependent on the degree of demethylation. The mutagenicity of the presumptive primary metabolite, hydroxymethyl pentamethylphosphoramide (HM-Me5-PA), is comparable to HMPA and can be decreased considerably by inhibition of the metabolism by 1-phenylimidazole or iproniazid. This suggests that further oxidative metabolism is required for mutagenic activity. The mutagenicity of the doubly hydroxylated HMPA metabolite, N,N''-bis(hydroxymethyl)-tetramethylphosphoramide (N,N''-(HN)2-Me4PA) can also be decreased by inhibition of metabolism, whereas the 3-fold hydroxylated N,N''-N"0(HM)3-Me3-PA is not affected by pretreatment with enzyme inhibitors, indicating that no further oxidative metabolism is required for its activation. A second hydroxylation on 1 dimethylamino group, forming N,N-(HM)2-Me4-PA, results in a drastic loss of mutagenic activity. further oxidation of HM-Me5-PA to formyl pentamethylphosphoramide (formyl--Me50PA) also leads to a strong reduction of the genotoxic activity. The rearrangement product of N-oxidation, N-((bis(dimethylamino)phosphinyl)-oxy)dimethylamine (HMPOA) is not mutagenic in Drosphila. The very low mutagenicity of hexaethylphosphoramide (Et6-PA) allowed us to study the mutagenicity of some ethyl-hydroxymethyl hybrid compounds. for the ethylated phosphoramides also the presence of only 1 hydroxymethyl group is insufficient for mutagenic activity, whereas the introduction of 2 or 3 hydroxymethyl groups resulted in considerable genotoxicity in the sex-linked recessive lethal (SLRL) test as well as in the ring-X loss test. It is concluded that the bioactivation of HMPA in Drosophila proceeds via multiple metabolic hydroxylations to form mult

关键词： Hydroxylation Hexamethylphosphoramide Drosophila melanogaster chrom chromosomes tested 3 d 3 days Et ethyl f feeding HEPA hexaethylphosphoric triamide HM hydroxymethyl HMPA hexamethylphosphoric triamide HMPOA N -(( bis ( dimethylamino ) phosphinyl ) oxy ) dimethylamine HMPT hexamethylphosphorus triamide I injection 1 lethals Me methyl N,N-DMB NN -dimethylbenzylamine PA phosphoric triamide prog total number of progency SLRL sex-linked recessive lethal tri-CIPDMT 1-(2,4,6-trichlorophenyl)-3, 3-dimethyltriazene

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ANALYSIS Of MOUTHPART MOVEMENTS DURING fEEDING Of fRANKLINIELLA-OCCIDENTALIS (PERGANDE) AND fRANKLINIELLA-SCHULTZEI TRYBOM (THYSANOPTERA, THRIPIDAE)

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INTERNATIONAL JOURNAL Of INSECT MORPHOLOGY & EMBRYOLOGY 1989年第2-3期18卷 161-171页

作者： HUNTER, WB ULLMAN, DE University of Hawaii at Manoa Department of Entomology 3050 Maile Way Gilmore Hall rm 310 Honolulu Hawaii 96822 USA

Scanning electron microscopy was used to elucidate the morphology and sequential movement of thrips feeding structures in the 2 species, frankliniella occidentalis and f. schultzei, (Thysanoptera : Thripidae). The mouthcone consists of paired paraglossae, and fringed labral pad, a single, apically fused mandible and a pair of interlocking maxillae. The maxillae are open apically and form a feeding tube. Ten pairs of sensory pegs of 3 distinct morphological types (sensilla basiconica with a cuticular collar, sensilla basiconica without a cuticular collar, and sensilla trichodea) were found on the paraglossae. The possible function of these sensory structures in host finding and choice are discussed. No structure for rasping the leaf''s surface was found on the mouthcone or the ventral surface of the insect. Live specimens were observed feeding through Parafilm on artificial media (sucrose solution) and lettuce leaf tissue. These observations support earlier findings that thrips feed by piercing leaf cells with the mandible and ingesting cell contents through the feeding tube formed by the maxillary stylets. Based on these findings, we suggest that thrips be classified as piercing-sucking rather than rasping-sucking insects.

关键词： morphology scanning electron microscopy sensory structures a mouthcone b left prothoracic leg c compound eye d antennae e maxillary palps f labial palps g sensory pegs h axonic sheaths i axons j paraglossae k labral pad 1 small palpi m long slender palpi n single mandibular stylet o paired maxillary stylets

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ROLE Of HEMIN IN THE INHIBITION Of MUTAGENIC ACTIVITY Of 3-AMINO-1-METHYL-5H-PYRIDO[4,3-B]INDOLE (TRP-P-2) AND OTHER AMINOAZAARENES

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MUTATION RESEARCH 1989年第2期213卷 217-226页

作者： ARIMOTO, S HAYATSU, H OKAYAMA UNIV FAC PHARMACEUT SCI OKAYAMA 700 JAPAN

The mechanism of inhibition by hemin of the mutagenic activities of food pyrolysate aminoazaarenes, particularly that of Trp-P-2 (3-amino-1-methyl-5H-pyrido[4,3-b]indole), was investigated. Hemin efficiently inhibited the metabolic activation by S9 of Trp-P-2, as demonstrated by high-performance liquid chromatographic analysis of the reaction mixtures in which Trp-2 had been treated with S9 in the presence or absence of hemin. N-Hydroxy-Trp-P-2, an activated form of Trp-P-2 having direct mutagenicity on Salmonella typhimurium TA98, undergoes spontaneous oxidative degradation in its aqueous solution, and the presence of hemin in the solution accelerated the degradation significantly. The presence of excess hemin with N-hydroxy-Trp-P-2 completely abolished the mutagenic activity of this mutagen towards Salmonella. A UV-visible spectroscopic study has suggested the formation of a complex between hemin and N-hydroxy-P-2/Trp-P-2. In support of this view, the fluorescence spectrum of a Trp-p-2 solution was quenched efficiently by the addition of hemin. These observation indicate that this complex formation plays a role in the observed multiple actions of hemin. Similar inhibitory actions of hemin on several other direct-acting aminoazaarene mutagens are also described, as well as the inhibition activities of protoporphyrin, chlorophyllin, biliverdin and bilirubin.

关键词： Trp-P-2 Aminoazaarenes Heterocyclic amines Hemin Porphyrins Trp-P-2(NHOH) 3-hydroxyamino-1-methyl-5 H -pyrido[4,3- b ]indole Trp-P-2 3-amino-1,4-dimethyl-5 H -pyrido[4,3- b ]indole Trp-P-1 3-amino-1,4-dimethyl-5 H -pyrido[4,3- b ]indole GLu-P-1(NHOH) 2-hydroxyamino-6-methyl-dipyrido[1,2-1: r ′,2′- d ]imidazole Glu-P-1 2-amino-6-methyl-dipyrido[1,2,- a : 3′,2′- d ]imidazole Glu-P-2 2-aminodipyrido[1,2- a : 3′,2′- d ]imidazole AαC 2-amino-9 H -pyrido[2,3,- b ]indole MeAαC 2-amino-3-methyl-9 H -pyrido[2,3,- b ]indole IQ 2-amino-3-methylimidazo[4,5- f ]quinoline MeIQ 2-amino-3,4-dimethylimidazo[4,5- f ]quinoxaline MEIQx 2-amino-3,8-dimethylidazo[4,5- f ]quinoline HPLC high-performance liquid chromatography

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EXCITATION-ENERGY TRANSfER IN PHYCOBILISOMES AT -196-DEGREES-C ISOLATED fROM THE CYANOBACTERIUM ANABAENA-VARIABILIS (M-3) - EVIDENCE fOR THE PLURAL TRANSfER PATHWAYS TO THE TERMINAL EMITTERS

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BIOCHIMICA ET BIOPHYSICA ACTA 1989年第2期973卷 153-162页

作者： MIMURO, M YAMAZAKI, I TAMAI, N KATOH, T NATL INST BASIC BIOL OKAZAKI AICHI 444 JAPAN INST MOLEC SCI OKAZAKI AICHI 444 JAPAN KYOTO UNIV FAC SCI DEPT BOT KYOTO 606 JAPAN

Excitation energy transfer in phycobilisomes at -***. C was studied by means of the time-resolved fluorescence spectroscopy in the picosecond time-range. Supplemental data were obtained from the allophycocyanin-core complex. When the phycobilisomes were excited at 580 nM, at least nine fluorescence components were resolved by the time-resolved spectra and deconvolution of those spectra. Energy transfer among these components is not straightforward. At the phycocyanin level, two transfer pathways are probable;one may be among the .beta.-155 chromophores, and another among .alpha.-84 chromophores along the long axis of the phycobilisome rods. The .beta.-84 chromophores in intermediate discs of the rods might function as an energy pool by a fast equilibrium between .alpha.-84 and .beta.-84 chromophores. The .beta.-84 chromophores in the trimer next to the core-complex is the energy donor to the core-complex. At allophycocyanin level, two pathways of the energy transfer were also found;one from f660 to f686 through the f673. This corresponds to the energy flow from .beta.-84 chromophore without linker to 18.3 kDa polypeptide and finally to the ''anchor'' polypeptide. The other pathway is from the f666 to f680, i.e., from the .beta.-84 with a linker polypeptide to the .alpha.-subunit of allophycocyanin B. Two independent pathways in the energy transfer shown in this study basically agree with the assembly model of the core components proposed by Glazer (cf. Biochim. Biophys. Acta 768 (1984) 29-51).

关键词： Energy transfer fluorescence Photosynthesis: Phycobilisome Picosecond spectroscopy: (Cyanobacterium) (A. variabilis (M-3)) α-APR α-subunit of allophycocyanin B PC phycocyanin APC allophycocyanin f 623 a representative expression of fluorescence component whose maximum is located at 623 nm fWHM full-width at half-maximum SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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STATE TRANSITIONS IN A PHYCOBILISOME-LESS MUTANT Of THE CYANOBACTERIUM SYNECHOCOCCUS SP PCC-7002

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BIOCHIMICA ET BIOPHYSICA ACTA 1989年第1期974卷 66-73页

作者： BRUCE, D BRIMBLE, S BRYANT, DA PENN STATE UNIV DEPT MOLEC & CELL BIOL UNIVERSITY PK PA 16802 USA

State transitions were investigated in the cyanobacterium Synechococcus sp. PCC 7002 in both wild-type cells and mutant cells lacking phycobilisomes. Preillumination in the presence of DCMU induced State 1 and dark-adaptation induced State 2 in both wild-type and mutant cells as determined by 77 K fluorescence emission spectroscopy. Light-induced transitions were observed in the wild-type after preferential excitation of phycocyanin (State 2) or preferential excitation of Chl a (State 1). Light-induced transitions were also observed in the phycobilisome=less mutant after preferential excitation of short-wavelength Chl a (State 2) or carotenoids and long-wavelength Chl a (State 1). We conclude that the mechanism of the light-state transition in cyanobacteria does not require the presence of the phycobilisome. Our results contradict proposed models for the state transition, which require phosphorylation of, and an active role for, the phycobilisome.

关键词： Cyanobacterium State transition Phycobilisome mutant ( Synechococcus sp. PCC 7002) Chl chlorophyll PS Photosystem LHC light-harvesting complex DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea f 695 fluorescence yield at 695 nm f 720 fluorescence yield at 720 nm

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ADENOSINE RECEPTORS LINKED TO ADENYLATE-CYCLASE ACTIVITY IN HUMAN NEURO-BLASTOMA CELLS - MODULATION DURING CELL-DIffERENTIATION

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NEUROSCIENCE 1989年第3期30卷 819-825页

作者： ABBRACCHIO, MP CATTABENI, f CLEMENTI, f SHER, E UNIV MILAN SCH PHARM INST PHARMACOL SCI I-20122 MILAN ITALY UNIV MILAN DEPT MED PHARMACOL I-20122 MILAN ITALY UNIV MILAN CNR CTR CYTOPHARMACOL I-20122 MILAN ITALY

In IMR32 neuroblastoma cells, the two adenosine receptor agonists N6-R-phenylisopropyladenosine and 5''-N-ethylcarboxamidoadenosine dose-dependently stimulated membrane adenylate cyclase activity with potencies consistent with the presence of adenosine receptors of the A2-subtype. The S enantiomer of N6-R-phenylisopropyladenosine induced a significantly lower stimulation of adenylate cyclase, accordingly to its lower ability to activate adenosine receptors. These effects were selectively counteracted by the adenosine receptor antagonist theophylline and, conversely, were not affected by the A1-adenosine receptor selective blocker 8-cyclopentyl-1,3-dipropylxanthine. No adenosine receptors belonging to the A1-subtype seem, therefore, to be present in this cell line, as also shown by the lack of inhibitory activity of N6-R-phenylisopropyladenosine on both basal and forskolin-stimulated adenylate cyclase activity. Activation of A2-receptors did not modify intracellular basal calcium levels, did not influence calcium influx through voltage-dependent calcium channels and did not modify calcium influx and redistribution induced by muscarinic receptor activation. Prolonged exposure of cells to either N6-R-phenylisopropyladenosine or 5''-N-ethylcarboxamidoadenosine was associated with a small but significant degree of morphological differentiation, comparable to that induced by dibutyryl cAMP, and therefore presumably related to the prolonged increase of intracellular cAMP levels elicited by the two adenosine agonists. After cellular differentiation induced with either dibutyryl cAMP or 5-bromodeoxyuridine, a selective desensitization of A2-receptor stimulated adenylate cyclase activity was found. IMR32 cells are a good model for studying the characteristics of the A2-adenosine receptors in a human neuronal tissue and its regulation during neuronal differentiation or pharmacological treatments.

关键词： ATP adenosine triphosphate 5-BrdU 5-bromodeoxyuridine cAMP cyclic adenosine-3',5'-monophosphate DMSO dimethylsulphoxide DPCPX 8-cyclopentyl-1,3-dipropylxanthine (PD-116,948) DTT dithiothreitol EGTA ethylene-glycol-bis(beta-amino-ethylether)- N , N , N ', N '-tetraacetic acid f − fluoride anion GTP guanosine 5'-triphosphate HEPES N -2-hydroxyethylpiperazine- N '-2-ethanesulphonic acid MEM minimum essential medium NECA 5'- N -ethyl-carboxamidoadenosine R-PIA N 6 - R -phenylisopropyl-adenosine S-PIA N 6 - S -phenylisopropyladenosine TH tyrosine hydroxylase

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THE PROGENITOR Of ATP SYNTHASES WAS CLOSELY RELATED TO THE CURRENT VACUOLAR H+-ATPASE

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fEBS LETTERS 1989年第1期247卷 147-153页

作者： NELSON, H NELSON, N ROCHE INST MOLEC BIOL ROCHE RES CTRNUTLEYNJ 07110

The gene encoding the proteolipid of the vacuolar H+-ATPase of yeast was cloned and sequenced. The deduced amino acid sequence of the yeast protein is highly homologous to that of the proteolipid from bovine chromaffin granules. In contrast to other membrane proteins the transmembrane segments of the bovine and yeast proteolipids were much more conserved than the hydrophilic parts. The fourth transmembrane segment, which contains the DCCD-binding site, was conserved 100%. Comparison of vacuolar and eubacterial proteolipids revealed a homology which pointed to a common ancestral gene that underwent gene duplication to form the vacuolar proteolipids. Additional support for this notion came from the amino acid sequences of subunits involved in the catalytic sectors of archaebacterial ATP synthase and plant and yeast vacuolar H+-ATPases, which reveal extensive sequence homology. Slight, but significant, homology between the archaebacterial and eubacterial ATP synthases was observed. These observations might suggest that the progenitor of ATP synthases was closely related to the present vacuolar H+-ATPases.

关键词： ATPase, vacuolar H + - ATP synthase Proteolipid Evolution f-ATPase, the eubacterial type H + -ATPase (f 0 f 1 ) V-ATPase, vacuolar type H + -ATPase DCCD, N , N ′-dycyclohexylcarbodiimide

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