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61.
Dominique Rumeau Stephan Cuiné Laurent Fina Nathalie Gault Michel Nicole Gilles Peltier 《Planta》1996,199(1):79-88
The intracellular compartmentation of carbonic anhydrase (CA; EC 4.2.1.1), an enzyme that catalyses the reversible hydration of CO2 to bicarbonate, has been investigated in potato (Solanum tuberosum L.) leaves. Although enzyme activity was mainly located in chloroplasts (87% of total cellular activity), significant activity (13%) was also found in the cytosol. The corresponding CA isoforms were purified either from chloroplasts or crude leaf extracts, respectively. The cytosolic isoenzyme has a molecular mass of 255 000 and is composed of eight identical subunits with an estimated M
r of 30000. The chloroplastic isoenzyme (M
r 220000) is also an octamer composed of two different subunits with M
r estimated at 27 000 and 27 500, respectively. The N-terminal amino acid sequences of both chloroplastic CA subunits demonstrated that they were identical except that the M
r-27 000 subunit was three amino acids shorter than that of the M
r-27 500 subunit. Cytosolic and chloroplastic CA isoenzymes were found to be similarly inhibited by monovalent anions (Cl–, I–, N
3
-
and NO
3
-
) and by sulfonamides (ethoxyzolamide and acetozolamide). Both CA isoforms were found to be dependent on a reducing agent such as cysteine or dithiothreitol in order to retain the catalytic activity, but 2-mercaptoethanol was found to be a potent inhibitor. A polyclonal antibody directed against a synthetic peptide corresponding to the N-terminal amino acid sequence of the chloroplastic CA monomers also recognized the cytosolic CA isoform. This antibody was used for immunocytolocalization experiments which confirmed the intracellular compartmentation of CA: within chloroplasts, CA is restricted to the stroma and appears randomly distributed in the cytosol.Abbreviations BSA
bovine serum albumin
- CA
carbonic anhydrase
- PMSF
phenylmethylsulphonyl fluoride
- BAM
benzamidine
- DTT
dithiothreitol
- 2-ME
2-mercaptoethanol
- PVDF
polyvinylidene difluoride
The authors thanks P. Carrier and Dr. B. Dimon for technical assistance with the mass-spectrometry measurements. 相似文献
62.
Insa Flechsler Annette G. Beck-Sickinger Holger Stephan Robert Sheppard Günther Jung 《Journal of peptide science》1995,1(3):191-200
Cleavage and kinetic studies have been carried out using commercially obtained H-Tyr(tBu)-5-(4′-aminomethyl-3′,5′-dimethoxyphenoxy)valeric acid-TentaGelS (H-Tyr(tBu)-4-ADPV-TentaGelS) and H-Tyr (tBu)-4-ADPV-Ala-aminomethyl-resin (H-Tyr(tBu)-4-ADPV-AM-resin) prepared from commercially available resin and loaded with commercially available Fmoc-4-ADPV-OH amide anchor. Cleavage with pure trifluoroacetic acid (TFA) gave the intermediate H-Tyr-4-ADPV-NH2, which was then degraded to H-Tyr-NH2, and cleavage with TFA/dichloromethane (1:9) yielded H-Tyr-4-ADPV-NH2 which could be isolated in preparative amounts. Cleavage reactions with 15N-labelled H-Ala-4-ADPV-[15N]-Gly-AM-resin yielded the intermediate H-Ala-4-ADPV-NH2, which contained no 15N as demonstrated by 1H-NMR. The analysis of the commercial Fmoc-4-ADPV-OH amide anchor showed the presence of Fmoc-4-ADPV-4-ADPV-OH as an impurity in high amounts. This dimeric anchor molecule is the cause of formation of the anchor-linked peptide intermediate obtained during the cleavage from the resin. The particularly high acid-lability of the amide bond between the two ADPV moieties was utilized to synthesize sidechain and C-terminally 4-ADPV protected pentagastrin on a double-anchor resin, and to cleave it using 5% trifluoroacetic acid in dichloromethane. This method may offer a new way for the synthesis of protected peptide amides with improved solubility to be used in fragment condensation. 相似文献
63.
Abstract. Altitudinal and latitudinal distribution limits of trees are mainly controlled by temperature. Therefore climate warming is expected to induce upslope or poleward migrations. In the Swiss Central Alps, summers in the period 1982-1991 were on average 0.8 °C warmer than those of the period 30 yr before. We investigated whether populations of conifers at the montane Pinus sylvestris-Pinus cembra ecocline exhibit demographic trends in response to that warming. We found no evidence for this. Young seedlings of Pinus sylvestris, the species which is expected to expand its range upward in a warmer climate, were virtually absent from all sites, whereas large fractions of Pinus cembra populations were observed in the seedling and juvenile categories even below the present lower distribution limit of adult trees. This suggests that there are no major altitudinal shifts in response to the recent sequence of warmer summers. Germination and seedling survival trials with Pinus sylvestris suggest that temperature per se would not exclude this species even from establishing at the current treeline in the Swiss Central Alps. Similar results were found at the polar treeline. Phytotron tests of seedling survival showed much less drought resistance in Pinus sylvestris than in Pinus cembra which is in contrast to their phytogeographic distributions. Thus, the montane pine ecocline in the Swiss Central Alps seems to be stabilized by species interactions and may not be directly responsive to moderate climatic change, which needs to be taken into account in predictive attempts. 相似文献
64.
During the first 8 days of germination the Ricinus seedling is supplied with all nutrients by the endosperm via phloem transport. In 4- to 8-days-old seedlings the concentrations and contents of Fe, Cu, Mn and Zn, and nicotianamine (NA) in the endosperm, cotyledons, hypocotyl and roots were estimated. From the data obtained translocation rates and flow profiles for the metals were established. The main sink for Fe, Mn and Zn were the cotyledons whereas Cu was mainly imported into the hypocotyl. Maximum flow rates occurred between days 5 and 7, for Zn between days 6 and 8.The time kinetics of NA and divalent metal ion concentrations and contents are interpreted as co-transport. The role of NA as transport vehicle of micronutrients in the sieve tubes is discussed. 相似文献
65.
Dipl. Geol. Jens Hefter Dipl. Geol. Volker Thiel Dr. Angela Jenisch Dr. Ursula Galling Dr. Stephan Kempe Dr. habil. Walter Michaelis 《Facies》1993,29(1):93-105
Summary Biomarker investigations were applied to the hydrocarbon fractions of three Recent (cyanobacterial mat, Lake Van microbialite
and Lake Satonda microbialite) and two Late Jurassic carbonate samples obtained from sponge bioherms. The relative concentrations
ofn-alkanes, monomethyl alkanes, acyclic isoprenoids, steroids and hopanoids in these samples are studied and their probable
biological precursors are discussed. Normal alkanes with carbon chain lengths ranging from C15 to C34 and monomethyl alkanes ranging from C17 to C21 with a varying methyl branching pattern are found. The major hydrocarbons are low molecular weight (LMW)n-alkanes (C15–C21) with a slight to strong predominance ofn-heptadecane (C17). High molecular weight (HMW)n-alkanes occur in low to moderate relative concentrations showing a preference of odd-carbon numbered compounds with a maximum
at C29. Within the acyclic isoprenoids, pristane, phytane/phytene, pentamethyl-eicosane, squalane and lycopane could be identified.
Polycyclic terpenoids of the sterane and/or hopane type are present in all carbonate samples. The carbon numbers of these
components range from 27 to 29 and 27 to 32, respectively. These organic compounds identified can be attributed to various
source organisms such as cyanobacteria, archaebacteria, algae and vascular plants. All hydrocarbon fractions of the samples
are characterized by moderate to high relative concentrations of compounds derived from cyanobacteria, signifying the role
of these organisms as contributors to the Recent as well as to the Late Jurassic carbonate deposits. 相似文献
66.
At altitudes between 1300 m to 2100 m in the Etna massif (Sicily), an endemic species of theBetula genus,Betula aetnensis Rafin, grows in a well-defined microclimatical context. Aboveground biomass and nutrient content studies within one stand revealed no significant differences from the otherBetula species, normally found in colder more temperate climate regions.Throughout the studied sites, biomass production, nutrient cycling and various structural or physiological characteristics (leaf area index) varied very little.Other researches indicate that the originality ofBetula aetnensis lies more in the histological or anatomical characteristics of its water conducting system which enables the species to adapt to Mediterranean-climate summer droughts in the Etna massif.
Riassunto Sull'Etna, tra 1300 e 2100 m d'altitudine, in una zona microclimaticamente ben definita del versante nordorientale, si rinviene laBetula aetnensis Rafin.Dallo studio della fitomassa e della mineralomassa aerea del bosco di Monte Baracca, è emerso che non vi sono differenze notevoli con le altre specie indagate del genereBetula, più caratteristiche dei climi temperati e freddi.La produzione di biomassa, cosi come la gestione degli elementi nutritivi, è molto simile ai diversi popolamenti già indagati, cosi come certe caratteistiche strutturali e fisiologiche (leaf area index).L'originalità dellaBetula aetnensis è da ricercarsi nel vantaggio che ne ricava, a livello endogeno, sfruttando le caratteristiche istologiche ed anatomiche del suo apparato conduttore, che le consentono un efficace ed eccellente adattamento alle condizioni di siccità estive particolari del clima mediterraneo del vulcano.相似文献
67.
The preparation and X-ray structure of [Ag(9-EtGH-N7)2]NO3·H2O(9-EtGH=neutral 9-ethylguanine) is reported. The compound crystallizes in the triclinic system, space group P
with a=7.063(6), b=7.153(3), c=11.306(10) Å, α=83.36(6), β=76.66(7), γ=81.44(6)°. The cation is centrosymmetric with Ag(I) coordinated via two N7 positions and Ag---N7 bond lengths of 2.11(1) Å. Applying 109Ag NMR spectroscopy, complex formation constants for both the 1:1 complex (log β1=0.6) and the title compound (log β2=1.6) in Me2SO have been determined. 相似文献
68.
H Owaki R Varma B Gillis J T Bruder U R Rapp L S Davis T D Geppert 《The EMBO journal》1993,12(11):4367-4373
69.
Hydrolysis of phosphatidylcholine couples Ras to activation of Raf protein kinase during mitogenic signal transduction. 总被引:12,自引:6,他引:6
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H Cai P Erhardt J Troppmair M T Diaz-Meco G Sithanandam U R Rapp J Moscat G M Cooper 《Molecular and cellular biology》1993,13(12):7645-7651
We have investigated the relationship between hydrolysis of phosphatidylcholine (PC) and activation of the Raf-1 protein kinase in Ras-mediated transduction of mitogenic signals. As previously reported, cotransfection of a PC-specific phospholipase C (PC-PLC) expression plasmid bypassed the block to cell proliferation resulting from expression of the dominant inhibitory mutant Ras N-17. In contrast, PC-PLC failed to bypass the inhibitory effect of dominant negative Raf mutants, suggesting that PC-PLC functions downstream of Ras but upstream of Raf. Consistent with this hypothesis, treatment of quiescent cells with exogenous PC-PLC induced Raf activation, even when normal Ras function was blocked by Ras N-17 expression. Further, activation of Raf in response to mitogenic growth factors was blocked by inhibition of endogenous PC-PLC. Taken together, these results indicate that hydrolysis of PC mediates Raf activation in response to mitogenic growth factors. 相似文献
70.