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1.
Summary Short oligocytidylates can act as templates for the self-condensation of guanosine 5-phosphorimidazolide. In the absence of a catalytic metal ion or in the presence of Pb2+ a noticeable template effect is already observed with the dimer and the yield of long oligomers reaches a plateau with a hexamer template. Short templates give oligomers longers than the template length. The products are predominantly 2-5 linked for the Pb2+-catalyzed reaction while mixed linkages are observed in the uncatalyzed reaction.In the presence of Zn2+, a template effect is first observed with the pentamer and is maximal by the heptamer. The products are predominantly 3-5 linked. Oligomers shorter than or as long as the template are obtained in substantial yield, and longer products in much lower yields.Abbreviations G
Guanosine
- Gp
guanosine 2(3)-phosphate
- pG
guanosine 5-phosphate
- Gp!
guanosine cyclic 2,3-phosphate
- ImpG
guanosine 5-phosphorimidazolide
- ImpG*
[8-14C]-guanosine 5-phosphorimidazolide
- pGp
5-phosphoguanosine 2(3)-phosphate
- G2pG
guanylyl-[2-5]-guanosine
- G3pG
guanylyl-[3-5]-guanosine
- ImpGpG
5-phosphorimidazolide of GpG
- (pG)n (n = 2,3)
oligomers of pG
- GppG
P1, P2-diguanosine 5-diphosphate
- GppGpG
5-[guanosine 5-pyrophosphate] of GpG
- NH2pG
guanosine 5-phosphoramidate
- (pG)4+
tetramer and higher oligoguanylates with 5 terminal phosphate
- oligo(G)
oligoguanylate
- Cp
cytidine 2(3)-phosphate
- Cp!
cytidine cyclic 2,3-phosphate
- (Cp)n–1 Cp! (n= 2,3,4)
oligocytidylates terminated by 5-OH groups and 2,3-cyclic phosphates
- oligo(C)
oligocytidylate
- poly(C)
polycytidylic acid
- poly(U)
polyuridylic acid
- poly(C,G)
random copolymer of C and G
- BAP
bacterial alkaline phosphatase (E. coli)
- EDTA
ethylenediaminetetraacetic acid
- Rf
chromatographic mobility 相似文献
2.
P. G. Kostyuk S. V. Viatchenko-Karpinskii M. B. Sedova V. I. Teslenko 《Neurophysiology》1994,26(6):331-333
Effects of different forms of C2-5A (2,5ApApA; 2,5ApApepoxyA; and 3,5ApApA) on high voltage-activated (HVA) calcium currents in GH3 cells were studied using the whole-cell patch-clamp recording technique. Addition of 10 µM 2,5ApApA, a core (dephosphorylated) oligoadenylate, to the pipette solution induced an increase in HVA calcium current. Ten minutes after the whole-cell configuration was established, the current magnitude was enhanced about twofold compared with that observed at 2 min. High concentration of Mg2+ (5 µM) in the pipette solution blocked this effect. 2,5ApA and 3,5ApApA oligoadenylates, and products of 2,5A hydrolysis, adenosine and AMP, did not change the value of HVA current. A chemically modified analog of 2,5ApApA (2,5ApApepoxyA) has been the oligoadenylate most stable under phosphodiesterase action. Addition of 2,5ApApepoxyA to the pipette solution under the same conditions caused a smaller effect than 2,5ApApA did.Neirofiziologiya/Neurophysiology, Vol. 26, No. 6, pp. 405–408, November–December, 1994. 相似文献
3.
The cyclic 35-nucleotide phosphodiesterase D3 was purified from Sinorhizobium fredii MAR-1. The native enzyme had a molecular weight of approximately 44.5kDa and a subunit molecular weight of approximately 21kDa as judged by SDS-gel electrophoresis. The pH optimum of the enzyme for the hydrolysis of cyclic AMP was approximately 6.0 with both acetate and Tris-maleate buffers. The optimum temperature for hydrolysing cyclic AMP was approximately 50C. No metal ion was required for activity and EDTA up to 2.5mM did not markedly affect the enzyme. However, methylxanthines, adenine and adenosine as well as 5-AMP, ATP, ADP and metal ions like Zn2+, Fe2+, Pb2+, Al3+ and Fe3+, were strongly inhibitory at 2.5mM.The D3 enzyme could hydrolyse both cyclic AMP and cyclic GMP with the apparent K
m for cyclic AMP of approximately 0.23M. 相似文献
4.
Wang Y 《Journal of biomolecular NMR》2004,30(3):233-244
For an amino acid in protein, its chemical shift, (, )s, is expressed as a function of its backbone torsion angles ( and ) and secondary state (s): (, )s=, )_coil+(, )_s, where (, )coil represents its chemical shift at coil state (s=coil); (, )s (s=sheet or helix) is herein defined as secondary structural effect correction factor, which are quantitatively determined from Residue-specific Secondary Structure Shielding Surface (RSS) for 13CO, 13C, 13C,1H, 15N, and 1HN nuclei. The secondary structural effect correction factors defined in this study differ from those in earlier investigations by separating out the backbone conformational effects. As a consequence, their magnitudes are significantly smaller than those earlier reported. The present (, )sheet and (, )helix were found varying little with backbone conformation and the 20 amino acids, specifically for 13CO, 13C, and 1H nuclei. This study also carries out some useful investigations on other chemical shift prediction approaches – the traditional shielding surfaces, SHIFTS, SHIFTX, PROSHIFT, and identifies some unexpected shortcomings with these methods. It provides some useful insights into understanding protein chemical shifts and suggests a new route to improving chemical shifts prediction. The RSS surfaces were incorporated into the program PRSI [Wang and Jardetzky, J. Biomol. NMR, 28: 327–340 (2004)], which is available for academic users at http://www.pronmr.com or by sending email to the author (yunjunwang@yahoo.com). 相似文献
5.
Summary Evaporation of a solution of thymidine plus either theexo or theendo diastereomer of uridine cyclic 2,3-O, O-phosphorothioate (U > p(S) in 1,2-diaminoethane hydrochloride buffer gave the 2,5 and 3,5 isomers of (P-thio) uridylylthymidine (Up(S)dT) in a ratio of 1:2 with a combined yield of about 20%. These isomers were re-converted to U > p(S) and dT by a reaction that is known to proceed by an in-line mechanism. Both the 2,5 and 3,5 isomers gave as product the same diastereomer of U > p(S) that had been used originally in their formation. These dry-state prebiotic reactions (Verlander, Lohrmann, and Orgel 1973) are thus shown to be stereospecific, and both the 2,5 and 3,5 internucleotide bonds are formed by an in-line mechanism.Abbreviations DAE
1,2-diaminoethane
- HPLC
high pressure liquid chromatography
- RNase
bovine pancreatic ribonuclease A, EC 3.1.4.22
- TEAB
triethylammonium bicarbonate
- tris
tris(hydroxymethyl)aminomethane
- UMP(S)
uridine monophosphorothioate
- U > p
uridine cyclic 2,3-phosphate
- U > p(S)
uridine cyclic 2,3-O, O-phosphorothioate
- Up(S)dT
(P-thio)uridylylthymidine
- U2p(Rp-S)5dT
(P-thio)uridylylthymidine with theR configuration at phosphorous, and a 2,5 internucleotide linkage 相似文献
6.
Silva E.M. Escamilla Dendooven L. Reynell J.A. Uscanga Ramírez A.I. Monroy Alatorre G. González de la Torre Martínez M. 《World journal of microbiology & biotechnology》1999,15(6):753-755
Strain H-984 of G. fujikuroi grown for 38h in a shake flask with medium containing 20g glucose l–1, 3g yeast extract l–1, 2.5g NH4NO3 l–1, 0.5g KH2PO4 l–1, 0.1g MgSO4 l–1, 1g CaCO3 l–1, and inoculated into a bioreactor with medium containing 60g glucose l–1; 1g NH4Cl l–1; 3g KH2PO4 l–1 and 1.5g MgSO4 l–1 produced 1100mg gibberellic acid l–1. 相似文献
7.
Summary We have studied the reactions between adenosine 5-phosphorimidazolide and 9-(2-amino-2-deoxyxylofuranosyl) adenine (I) or 3-methylamino-3-deoxyadenosine (II), both with and without a poly (U) template. We find that both amino compounds react much more rapidly than does adenosine, in the absence of a template. The rate of reaction is greatly enhanced by a poly (U) template in the case of I, but the enhancement is slight in the case of II.Abbreviations A
adenosine
- xylo ANH2
9-(2-amino-2-deoxy--D-xylofuranosyl) adenine
- ANHMe
3-methylamino-3-deoxyadenosine
- ImpA
adenosine 5-phosphorimidazolide
- A3 pA
adenylyl-[35]-adenosine
- A2 pA
adenylyl-[25]-adenosine
- UNPA
adenylyl-[52]-2-amino-2-deoxyuridine
- xylo ANPA
9-[adenylyl-(52)-2-amino-2-deoxy--D-xylofuranosyl]adenine
- A(NMe)pA
adenylyl-[53]-3-methylamino-3-deoxyadenosine
- pA
adenosine 5phosphate
- AppA
P1, P2-diadenosine 5pyrophosphate
- (pA)n
n = 2, 3 [2-5]-linked oligomers of pA
- A2 pA2 pA
[2-5]-linked trinucleoside diphosphate of A
- poly (U)
polyuridylic acid 相似文献
8.
Electron transport-linked proton translocation at nitrite reduction inCampylobacter sputorum subspeciesbubulus 总被引:1,自引:0,他引:1
Wytske de Vries H. G. D. Niekus Hugerien van Berchum A. H. Stouthamer 《Archives of microbiology》1982,131(2):132-139
Campylobacter sputorum subspeciesbubulus contains a membrane-bound nitrite reductase which catalyses the six-electron reduction of nitrite to ammonia. Formate andL-lactate are used as hydrogen donors. Cells ofC. sputorum grown with nitrate or nitrite contain cytochromes of theb-andc-type and a carbon monoxide-binding cytochromec. In addition, a special membrane-bound carbon monoxide-binding pigment is found. Nitrite reduction with formate orL-lactate as a hydrogen donor is strongly inhibited by 2-n-heptyl-4-hydroxyquinoline-N-oxide (HQNO). Nitrite reduction by bacterial suspensions with lactate as a hydrogen donor is strongly inhibited by carbonylcyanide-m-chlorophenyl-hydrazone (CCCP) whereas nitrite reduction with formate as a hydrogen donor is not inhibited at all. H+/O values and H+/NO
2
-
values were measured with ascorbate + N,N,N,N-tetramethyl-p-phenylenediamine (TMPD), formate (in the absence and presence of carbonic anhydrase) andL-lactate as a hydrogen donor. The results are summarized in a scheme for electron transport from formate or lactate to oxygen or nitrite which shows a periplasmic orientation of formate dehydrogenase and nitrite reductase and a cytoplasmic orientation of lactate dehydrogenase and oxygen reduction, and which shows proton translocation with a H+/2e value of 2.0. The H+/O and H+/NO
2
-
values predicted by this scheme are in good agreement with the experimental values.Abbreviations CCCP
carbonylcyanide-m-chlorophenylhydrazone
- HQNO
2-n-heptyl-4-hydroxyquinoline-N-oxide
- MTPP+
methyltriphenylphosphonium cation
- TMPD
N,N,N,N-tetramethyl-p-phenylenediamine; H+/O (H+/NO
2
-
), number of protons liberated in the outer bulk phase at the reduction of one atom O (one ion NO
2
-
); H+/2e (q+/2e), number of protons (charges) translocated across the cytoplasmic membrane during flow of two electrons to an acceptor 相似文献
9.
Masao Kawai Tatsuo Yamamoto Keiichi Yamada Masashi Yamaguchi Shigehiro Kurobe Hatsuo Yamamura Shuki Araki Yasuo Butsugan Kyoko Kobayashi Ryoichi Katakai Kazuki Saito Terumi Nakajima 《Letters in Peptide Science》1998,5(1):5-12
Derivatives of gramicidin S (GS) and its mono- and di-d-cyclohexylalanine (d-Cha) analogs possessing various protecting groups on Orn side chains were prepared. 1H NMR spectra of the unsymmetrically protected analogs [Orn(X)2,Orn(X)2,d-Cha4]GS were similar to the composites of the spectra of the symmetrical derivatives [Orn(X)2,2,d-Cha4,4]GS and [Orn(X)2,2]GS, revealing the proximity of the protecting groups of NH of Orn residues at the 2 and 2 positions to the side chains of d-Phe (or d-Cha) residues at the 4 and 4 positions, respectively. The results indicated the presence of H-bonds between the NH of Orn and the carbonyl of d-Phe residues in the i i + 2 sense and not in i i – 3, which was also supported by the ROESY analysis. The substantially strong H-bonds can explain the observed resistance of the urethane NH of the Orn side chains in the GS derivatives to the N-methylation with CH3I–Ag2O in DMF. 相似文献
10.
Conversion of methanol to CH4 has a large isotope effect so that a small contribution of methanol-dependent CH4 production may decrease the 13CH4 of total CH4 production. Therefore, we investigated the role of methanol for CH4 production. Methanol was not detectable above 10 M in anoxic methanogenic rice field soil. Nevertheless, addition of 13C-labeled methanol (99% enriched) resulted in immediate accumulation of 13CH4. Addition of 0.1 M 13C-methanol resulted in increase of the 13CH4 from –47 to –6 within 2 h, followed by a slow decrease. Addition of 1 M 13C-methanol increased 13CH4 to +500 within 4 h, whereas 10 M increased 13CH4 to +2500 and continued to increase. These results indicate that the methanol concentrations in situ, which diluted the 13C-methanol added, were 0.1 M and that the turnover of methanol contributed only about 2% to total CH4 production at 0.1 M. However, contribution increased up to 5 and 17% when 1 and 10 M methanol were added, respectively. Anoxic rice soil that was incubated at different temperatures between 10 and 37 °C exhibited maximally 2–6% methanol-dependent methanogenesis about 1–2 h after addition of 1 M 13C-methanol. Only at 50 °C, contribution of methanol to CH4 production reached a maximum of 10%. After longer (7–10 h) incubation, however, contribution generally was only 2–4%. Methanol accumulated in the soil when CH4 production was inhibited by chloroform. However, the accumulated methanol accounted for only up to 0.7 and 1.2% of total CH4 production at 37 and 50 °C, respectively. Collectively, our results show that methanol-dependent methanogenesis was operating in anoxic rice field soil but contributed only marginally to total CH4 production and the isotope effect observed at both low and high temperature. 相似文献
11.
Alternating poly(Glu-Leu) exhibits a random coilstructure in pure water at neutral pH. The addition of 0.5 equiv of Ca2+ induces a coil-to--sheet transitionand the addition of 0.15 equivof Fe3+ induces a coil-to--helixtransition. Conformational competition between thesetwo structures was studied by mixing preformed-sheets and -helices in differentproportions. Circular dichroism spectra clearly show that-sheets are favored at the expense of-helices in -sheet-rich mixtures. 相似文献
12.
Melanie S. Johnson James Simpson David J. MacEwan Angela Ison Roger A. Clegg Kevin Connor Rory Mitchell 《Molecular and cellular biochemistry》1995,146(2):127-137
The characteristics of PKC activation induced by a number of compounds were investigated using PKCs, partially-purified from sources with a naturally high abundance of certain Ca2+ dependent PKC isoforms. Native isoforms were used rather than PKC isoforms expressed from a baculovirus system to assess the effect of tissue specific factors on activity. However, some data using recombinant PKC were included for comparison.The presence of specific PKC isoforms in different tissues was determined using Western blot analysis. Protein kinase C , 1, , , and / were all present in rat midbrain cytosolic extract, PKC , 1, , and / were present in spleen cytosol, and PKC and / were present in COS 7 cell cytosol. The predominance of and activities in COS 7 and spleen extracts respectively was confirmed by enzymic assay.The PKC activity assay was configured such that the Ca2+ dependence of the PKC activity induced by different PKC activators could be determined. Phorbol 12,13-dibutyrate (PDBu) was virtually equipotent on the Ca2+-dependent PKC activity from midbrain and spleen and slightly less potent on that from COS 7 cells. In the absence of Ca2+, PDBu was considerably less potent overall (as, indeed, were the other PKC activators) and was less potent on COS 7 cell PKC than on those from midbrain or spleen. Mezerein was more potent than PDBu at inducing PKC activity in COS 7 cell extracts in either the absence or presence of Ca2+ whereas in the presence of Ca2+, mezerein was slightly less potent on midbrain and spleen than PDBu and equipotent in the absence of Ca2+. Maximum values for Ca2+-independent activation by mezerein indicated that this activator was particularly effective in recruiting Ca2+-dependent PKC isoform activity in a Ca2+ free environment. The greater potency of mezerein on PKC was confirmed using PKC and further purified from rat spleen by hydroxylapatite (HAP) chromatography. The effects of both PDBu and mezerein were investigated using anterior pituitary tissue where a particularly high potency of mezerein in the absence of Ca2+ was noted. The diacylglycerol, 1,2-dioctanoyl-sn-glycerol (DOG), appeared to cause little or no activation of native Ca2+-dependent isoforms in Ca2+ free conditions unlike another longer chain diacylglycerol, 1,2-dioleoyl-sn-glycerol. Also DOG activated midbrain PKCs more potently than PKCs from spleen or COS 7 cells (or lung and pituitary tissue) in the presence of Ca2+. The concentration-dependence of DOG was examined on PKC and PKC further purified from brain by HAP chromatography, revealing that DOG was equally potent on both of these isoforms derived from brain and on recombinant PKC . However, [3H]PDBu binding data using PKC purified from several sources gave very different IC50 values when DOG was used as a displacer, and in general these values correlated with the EC50 values recorded from the activity assay.The data presented here indicate that there are distinct differences in the activator pharmacology of different native PKC isoforms and between the same isoform expressed in different tissues, either because of post-translational modifications or some other tissue specific factor. 相似文献
13.
A quantitative analysis of JPH scalar couplings in nucleic acids is difficult due to small couplings to phosphorus, the extreme overlap of the sugar protons and the fast relaxation of the spins involved in the magnetization transfer. Here we present a new methodology that relies on heteronuclear Constant Time Correlation Spectroscopy (CT-COSY). The three vicinal 3JPH3, 3JPH5 and 3JPH5 scalar couplings can be obtained by monitoring the intensity decay of the Pi-H3i – 1 peak as a function of the constant time T in a 2D correlation map. The advantage of the new method resides in the possibility of measuring the two 3JPH5 and 3JPH5 scalar couplings even in the presence of overlapped H5/H5 resonances, since the quantitative information is extracted from the intensity decay of the P-H3 peak. Moreover, the relaxation of the H3 proton is considerably slower than that of the H5/H5 geminal protons and the commonly populated conformations of the phosphate backbone are associated with large 3JPH3 couplings and relatively small 3JPH5 / H5. These two facts lead to optimal signal-to-noise ratio for the P-H3 correlation compared to the P-H5/H5 correlation.The heteronuclear CT-COSY experiment is suitable for oligonucleotides in the 10–15 kDa molecular mass range and has been applied to the 30mer HIV-2 TAR RNA. The methodology presented here can be used to measure P-H dipolar couplings (DPH) as well. We will present qualitative results for the measurement of P-Hbase and P-H2 dipolar couplings in the HIV-2 TAR RNA and will discuss the reasons that so far precluded the quantification of the DPHs for the 30mer RNA. 相似文献
14.
James P. Ferris Gözen Ertem Vipin Agarwal 《Origins of life and evolution of the biosphere》1989,19(2):165-178
The reaction of the 5-AMP with water soluble carbodiimide (EDAC) in the presence of Na+-montmorillonite 22A results in the formation of 2,5-(pA)2 (18.9%), 3,5-(pA)2 (11%), and AppA (4.8%). When poly(U) is used in place of the clay the product yields are 2,5-(pA)2 (15.5%), 3,5-(pA)2 (3.7%) and AppA (14.9%). The 3,5-cyclic dinucleotide, 3,5-c(pA)2, is also formed when poly(U) is used. AppA is the principal reaction product when neither clay nor poly(U) is present in the reaction mixture. Products which contain the phophodiester bond are formed at different ionic strengths, pH and temperatures using Na+-montmorillonite. Phosphodiester bond formation was not observed when Cu2+-montmorillonite was used or when DISN was used in the place of EDAC. The extent catalysis of phophodiester bond formation varied with the particular clay mineral used. Those Na+-clays which bind 5-AMP more strongly are better catalysts. Cu2+-montmorillonite, which binds 5-AMP strongly, exhibits no catalytic activity. 相似文献
15.
The Ca2+-binding properties of calmodulin purified from zucchini (Cucurbita pepo L.) has been determined. A value of 3.3 mol Ca2+ per mol of zucchini calmodulin was measured at pH 7.5 by equilibrium chromatography. The far-and near-UV circular-dichroic spectra of the Ca2+-and Mg2+-saturated as well as from the metal-free forms of zucchini calmodulin reveal that upon Ca2+-binding the -helix content increases. A comparison with the spectra of vertebrate calmodulin indicates that both calmodulin have a similar secondary structure, similar Ca2+-induced conformational changes and the same number of Ca2+-binding sites.Abbreviations CAPP
10-(3-aminopropyl)-2-chloro-phenothiazine
- EGTA
ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid
- EDTA
ethylenediaminetetraacetic acid
Dedicated to Prof. Dr. Karl Decker on the occasion of his 60th birthday 相似文献
16.
Uptake kinetics of nitrogen derived from sewage–seawater mixtures (2.5–20% v/v effluent) were determined in the laboratory for Ulva rigida (Chlorophyceae) native from Bahía Nueva (Golfo Nuevo, Patagonia, Argentine). In terms of nitrogen concentration, experimental enrichment levels varied between 53.7 and 362.3M of ammonium and between 0.77 and 6.21M of nitrate+nitrite. Uptake rates were fitted to the Michaelis–Menten equation, with the following kinetic parameters: ammonium: Vmax = 591.2molg–1h–1,
K
s=262.3M, nitrate+nitrite: V
max=12.9molg–1h–1, K
s=3.5M). Both nutrients were taken up simultaneously, but ammonium incorporation was faster in all cases. The results show a high capability of Ulva rigida to remove sewage-derived nitrogen from culture media. In the field, most of the nitrogen provided by the effluent would be tied up in algal biomass, supporting low nitrogen levels found at a short distance away from the source. 相似文献
17.
We examined the effects of changing KCl concentration on the secondary structures of -actinins using circular dichroism (CD), 1,1-bis(4-anilino) naphthalene-5,5-disulfonic acid (bisANS) fluorescence and proteolysis experiments. Under near-physiological conditions, divalent cations also were added and changes in conformation were investigated. In 25 mm KH2PO4, pH 7.5, increasing KCl from 0 to 120 mm led to decreases in -helix conformation for brain, platelet and heart -actinins (40.5-30.2%, 65.5-37.8% and 37.5-27.8%, respectively). In buffered 120 mm KCl, 0.65 mm calcium produced small changes in the CD spectra of both brain and platelet -actinin, but had no effect on heart -actinin. bisANS fluorescence of all three -actinins also showed significant changes in conformation with increasing KCl. However, in buffered 120 mm KCl increasing concentrations of Ca2+ or Mg2+ did not have significant effects on the bisANS fluorescence of any -actinin. Digestion of brain, platelet and heart -actinins with -chymotrypsin showed an increase of proteolytic susceptibility in 120 mm KCl. These experiments also showed that increasing the concentration of Ca2+ or Mg2+ led to greater changes in digestion fragment patterns in the absence of KCl than in the presence of 120 mm KCl. The results suggest that -actinins exist in different conformations depending on the ionic strength of the medium, which could explain the differences in calcium and F-actin binding results obtained from different -actinins. 相似文献
18.
Summary We examined the ionic regulation of tip growth inNeurospora crassa by a combination of electrophysiology and confocal microscopy. To determine if transmembrane ionic fluxes are required for tip growth, we voltage clamped the membrane from –200 to +50 mV. In this voltage range, transmembrane ionic fluxes would either reverse (e.g., K+) or change dramatically (e.g., Ca2+ influx) but had no effect on hyphal growth rates. Therefore, ionic fluxes (including Ca2+ influx) may not be required for tip growth. However, intracellular Ca2+ may still play an obligatory role in tip growth. To assess this possibility, we first increased cytosolic Ca2+ directly by ionophoresis. Elevated Ca2+ induced subapical branch initiation, often multiple tips. At hyphal tips, fluorescence ratio imaging using fluo-3 and fura-red revealed a pronounced tip-high Ca2+ gradient within 10 m of the tip in growing hyphae which was not observed in nongrowing hyphae. Injection of the Ca2+ chelator 1,2-bis(ortho-aminophenoxy)ethane-N,N,N,N-tetrapotassium acetate consistently inhibited growth concomitantly with a depletion of intracellular Ca2+ and dissipation of the tip-high gradient. We conclude that Ca2+ plays a regulatory role in tip initiation and the maintenance of tip growth. Because plasma membrane ionic fluxes do not play a role in tip growth, we suggest that the tip-high Ca2+ gradient is generated from intracellular Ca2+ stores in the ascomyceteN. crassa.Abbreviations BAPTA
1,2-bis(ortho-aminophenoxy)ethane-N,N,N,N-tetrapotassium acetate
- [Ca2+]i
intracellular Ca2+ concentration
- fluo-3
2,7-dichloro-6-hydroxy-3-oxo-9-xanthenyl-4-methyl-2,2-(ethylenedioxy)dianiline-N,N,N,N-tetraacetic acid 相似文献
19.
Crude extracts or supernatants of broken cells of Clostridium formicoaceticum reduce unbranched, branched, saturated and unsaturated carboxylates at the expense of carbon monoxide to the corresponding alcohols. The presence of viologens with redox potentials varying from E
0=-295 to-650 mV decreased the rate of propionate reduction. The more the propionate reduction was diminished the more formate was formed from carbon monoxide. The lowest propionate reduction and highest formate formation was observed with methylviologen. The carbon-carbon double bond of E-2-methyl-butenoate was only hydrogenated when a viologen was present. Formate as electron donor led only in the presence of viologens to the formation of propanol from propionate. The reduction of propionate at the expense of a reduced viologen can be followed in cuvettes. With respect to propionate Michaelis Menten behavior was observed. Experiments are described which lead to the assumption that the carboxylates are reduced in a non-activated form. That would be new type of biological reduction.Non-standard abbreviations glc
Gas liquid chromatography
- HPLC
high performance liquid chromatography
- RP
reverse phase; Mediators (the figures in parenthesis of the mediators are redox potentials E
0 in mV)
- CAV2+
carbamoylmethylviologen, 1,1-carbamoyl-4,4-dipyridinium dication (E
0=-296 mV)
- BV2+
benzylviologen, 1,1-dibenzyl-4,4-dipyridinium dication (E
0=-360 mV)
- MV
methylviologen, 1,1-dimethyl-4,4-dipyridinium-dication (E
0=-444 mV)
- DMDQ2+
dimethyldiquat, 4,4-dimethyl-2,2-dipyridino-1,1-ethylendication (E
0=-514 mV)
- TMV2+
tetramethylviologen, 1,1,4,4-tetramethyl-4,4-dipyridinium dication (E
0=-550 mV)
- PDQ2+
propyldiquat, 2,2-dipyridino-1,1-propenyl dication (E
0=-550 mV)
- DMPDQ2+
dimethylpropyldiquat, 4,4-dimethyl-2,2-dipyridino-1,1-propenyl dication (E
0=-656 mV)
- PN
productivity number=mmol product (obtained by the uptake of one pair of electrons) x (biocatalyst (dry weight) kg)-1×h-1 相似文献
20.
Boris F. Minaev 《Molecular Engineering》1996,6(3):261-279
Recombination of the triplet state radical pair consisting of two hydrogen atoms catalysed by molecular oxygen is considered as a simulating example of a paramagnetic-exchange catalytic process. Intermolecular exchange interaction in the collision complex between the H2 and O2 molecules is calculatedab initio in STO-6G and 6–31 G* basis sets with complete active space configuration interaction. Calculations are done at a fixed O–H distance (3 Å), scanning the H–H bond length from 0.6 till 12 Å at the linear geometry of collision. The mixture of the triplet (T)3
u
+
and singlet (S)1
g
+
states of the hydrogen moiety is possible because both states have the same triplet symmetry in the collision complex with O2 (3
g
–
). A strong mixture of the S (1
g
+
, H2 +3
g
–
, O2) and T) and T (3
u
+
, H2 +3
g
–
, O2) states is actually obtained even at large H–H distances. The quintet and singlet states5,1(3
u
+
, H2 +3
g
–
, O2) are also considered for comparison of the exchange potentials. Atr(H–H)4.4 Å the S-T splitting is approximately constant (12 cm-1 in the STO-6G basis set; 55.5 cm-1 in the 6–31 G* basis set) and is determined by the exchange interaction between O2 and the nearest hydrogen atom in the O–O...H fragment. The paramagnetic catalyst can accelerate radical recombination through the triplet-singlet nonadiabatic transition to the lowest S reactive state when the radical encounter takes place in the vicinity of the catalyst. Though we do not consider the radical dynamics in a real solvent, which modulates the exchange potentials and the T-S transitions, the nature of this mechanism of spin catalysis is obvious. The electric polarization and charge transfer are important in the analysis of the exchange interaction and radical recombination potentials for all multiplets. In accordance with the concept of spin catalysis, the electronic spin-uncoupling mechanism, induced by O2 perturbation, has the same nature as other known catalytic processes of paramagnetic-exchange type. 相似文献