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1.
2D NMR spectroscopy and J coupling constant analysis are applied to resolve the structure of two photoproducts of thymidylyl-(35)-thymidine. These products are cyclobutane type thymine dimers possessing the cis-syn (the predominant one) and trans-syn geometry. The cis-syn is formed in an ANTI-ANTI conformation about the N-glycosyl linkages and resembles the normal base-stacked configuration. The glycosidic conformation in solution of the 5 terminal fragment differs from the crystal in which the less common SYN conformation is observed. In this isomer only the sugar pucker of the 3 terminal fragment is changed substantially with respect to the dinucleotide. The trans-syn isomer is formed in a SYN-ANTI glycosidic conformation. In this isomer the sugar puckers of both deoxyribose rings are affected and a preference for a pure 2-endo conformation is observed.Abbreviations dTpdT
2-deoxythymidylyl-(35)-2-deoxythymidine
- dTp[]dT
cyclobutane type photodimers of dTpdT
- dTp- and dTp[]-
their 5' terminal fragments (fragment A)
- -pdT and-[]pdT
their 3 terminal fragments (fragment B)
- RP-HPLC
reversed-phase high-performance liquid chromatography
- COSY
two-dimensional correlated spectroscopy
- 2D NOE
two-dimensional nuclear Overhauser spectroscopy 相似文献
2.
Summary We have synthesized 2(3)-O-(glycyl)-adenosine-5-(O-methylphos-phate), an analogue of the 3-terminus of aminoacylated tRNA. A 0.4M solution of this compound maintained at pH 8.2, yields 5.5% of diglycine and 11.5% of diketopiperazine, in addition to the hydrolysis products glycine and adenosine-5-(O-methylphosphate). Under the same conditions, glycine ethyl ester reacts much more slowly, but ultimately gives similar yields of diglycine and diketopiperazine.The aminolysis of 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) by free glycine is relatively inefficient, but serine reacts 20 times more rapidly and yields up to 50% of N-glycylserine. The prebiotic significance of these reactions is discussed.Abbreviations MepA
adenosine-5-(O-methylphosphate)
- MepA-gly
2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate)
- MepA-bis-gly
2,3-O-(bis-glycyl)-adenosine-5-(O-methylphosphate)
- DKP
diketopiperazine
- gly Et
glycine ethyl ester
- gly-ser
N-glycylserine
- O-gly-ser
O-glycylserine
- O-(gly)-gly-ser
O-(glycyl)-glycylserine
- Boc-gly
N-tert-butyloxycarbonylglycine
- MepA-Boc-gly
2(3)-O-(Boc-glycyl)-adenosine-5-(O-methylphosphate)
- MepA-bis-Boc-gly
2,3-O-(bis-Boc-glycyl)-adenosine-5(O-methylphosphate)
- (gly)2
diglycine
- (gly)3
triglycine 相似文献
3.
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
4.
Isopropylidenation of lactose with 2,2-dimethoxypropane in the presence ofp-toluenesulfonic acid gave two products, which were identified by1H- and13C-NMR as 2,35,63,4-tri-O-isopropylidenelactose dimethyl acetal (1) and its 6-O-(2-methoxy)-isopropyl derivative (2). These products were used for the synthesis of 2-O-methyllactose (7), 2,6-di-O-methyllactose (9) and 2-O-benzyllactose (13). 相似文献
5.
Lai-Xi Wang Nadejda V. Pavlova Su-Chen Li Yu-Teh Li Yuan C. Lee 《Glycoconjugate journal》1996,13(3):359-365
4-Methylumbelliferyl 6-O-benzyl--d-lactoside (6Bn-MU-Lac) and some related compounds were synthesizedvia different selective reactions including phase-transfer glycosylation. Their suitability as substrates for a fluorometric assay of ceramide glycanase (CGase) was evaluated. Among others, the 6Bn-MU-Lac, which is resistant to exogalactosidase, was found to be a suitable substrate for routine assay of the CGase activity. For American leech CGase, theK
m value is 0.232 mM at pH 5.
Abbreviations: CGase, ceramide glycanase; Gal, galactose; Glc, Glucose; Lac, lactose; MU, 4-methylumbelliferone; MU-Lac, 4-methylumbelliferyl -d-lactoside; bBn-Lac, 6-O-benzyl-lactose; 6Bn-MU-Lac, 4-methylumbelliferyl 6-Obenzyl--d-lactoside; 46Bd-MU-Lac, 4-methylumbelliferyl 4,6-O-benzylidene--d-lactoside; MU-Cel, 4-methylumbellifery -d-cellobioside; 46Bd-MU-Cel, 4-methylumbelliferyl 4,6-O-benzylidene--d-cellobioside; TLC, thin layer chromatography;1H-NMR, proton nuclear magnetic resonance; GSL, glycosphingolipids; CSA, 10-camphorsulfonic acid. See Scheme 1 for chemical structures. 相似文献
6.
Wei Tong Wang Fumito Matsuura Hernan Nunez Norman Ledonne Jr. Betty Baltzer Charles C Sweeley 《Glycoconjugate journal》1984,1(1):17-35
Ten previously unreported oligosaccharides have been purified from the urines of human subjects using a combination of gel filtration, ion exchange, and thin-layer chromatographies. Their structures were determined by direct probe mass spectrometry, methylation analysis, and proton NMR spectroscopy of the permethylated oligosaccharide alditols.On the basis of composition, the oligosaccharides could be divided into three groups. Five oligosaccharides containing glycerol were characterized as glucosyl1-1glycerol; glucosyl1-1glycerol; galactosyl1-1glycerol; glucosyl-1-1(fucosyl-1-2)glycerol and/or fucosyl-1-1(glucosyl-1-2)glycerol; and glucosyl-1-1(galactosyl-1-2)glycerol or galactosyl-1-1(glucosyl-1-2)glycerol. Four inositol-containing oligosaccharides were characterized as galactosyl1 (fucosyl1)inositol,N-acetylgalactosaminyl1 (fucosyl1)inositol, fucosyl1-2galactosyl1 (N-acetylgalactosaminyl1)inositol and fucosyl1-2galactosyl1-4-N-acetylglucosaminyl1(N-acetylgalactosaminyl1)inositol. Finally, galactosyl1-3(fucosyl1-2)galactosyl1-6galactosyl1-4(fucosyl1-3)glucose, an oligosaccharide with glucose at its reducing end, was tentatively identified. The significance and possible origins of the carbohydrate structures are discussed. 相似文献
7.
Purified myelin from fresh calf brain white matter was subfractionated in a discontinuous sucrose gradient; significant recovery of protein and 2,3-cyclic nucleotide 3-phosphohydrolase (CNP) and 5-nucleotidase (5N) activities occurred in all three obtained subfractions, the highest recovery being in the light subfraction; highest 5N and CNP specific activities were in medium myelin. Purified myelin was also subfractionated in a continuous sucrose gradient, with a similar localization of protein; CNP activity and 5N activity maxima suggest that myelin may be a predominant locus of 5N in bovine brain white matter. Freezing of brain white matter caused an increase in protein and in CNP and 5N total activity recoveries in denser myelin subfractions. Cytochemistry showed the reaction product of 5N in the whole myelin fraction to be associated with the innermost, outermost and medial compact myelin layers. Effects of non-ionic detergent (Lubrol WX) on 5N activity were studied, and the results also suggest the intrinsic nature of 5N as an ectoenzyme in myelin membranes. Lubrol WX was viewed as an advisable detergent for the stimulation of myelin 5N activity, but not for the solubilization of this enzyme. 相似文献
8.
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
9.
Inhibition of gap junction-mediated cell-cell communication might be a mechanism for several types of cellular dysfunctions, including tumor promotion. Although many different assays have been designed to measure gap junction-mediated intercellular communication, we applied a new technique, termed Fluorescence Redistribution After Photobleaching (FRAP), to assess the ability of a known tumor promoter, 2,2, 4,4, 5,5-hexabromobiphenyl (245-HBB), to inhibit cell-cell communication in a concentration-dependent manner. WB-F344 (rat epithelial) cells were plated at low density, exposed to noncytotoxic concentrations of 1, 5, or 20 µg 245-HBB/ml medium, and stained with 6-carboxyfluorescein diacetate. Single cells in pairs or clusters of touching cells in each exposure group were examined with FRAP. The results revealed an inverse correlation between the degree offluorescence redistribution in photobleached cells and the concentration of 245-HBB. Therefore, FRAP appears to be a sensitive and rapid technique for determining complete or partial inhibition of chemically induced intercellular communication in vitro. These results also provide further evidence for the ability of 245-HBB to inhibit gap junction-mediated cell-cell communication in a concentration-dependent manner.Abbreviations 6-CFDA
6-carboxyfluorescein diacetate
- FRAP
fluorescence redistribution after photobleaching
- 245-HBB
2,2, 4,4, 5,5-hexabromobiphenyl
Michigan Agricultural Experiment Station journal article No. 12531. 相似文献
10.
Adventitious shoot regeneration was compared among leaf, stem and petal explants of carnation (Dianthus caryophyllus L.) cv. Scania on MS medium containing different concentrations of 6-benzyladenine (BA) and -naphthaleneacetic acid (NAA). High frequency regeneration was obtained only from petal explants on the media containing 5 to 10 M BA with or without 5 M NAA. Among the cytokinins tested, N-2-chloro-4-pyridyl-N-phenylurea and N-1,2,3-thiadiazol-5-yl-N-N-phenylurea were more effective than BA, kinetin, N6-2-isopentenyl adenine and zeatin on regeneration from petal explants. Although, high frequency shoot regeneration was obtained from all petal explants harvested from various developmental stages of buds, a significant decrease in regeneration capacity was observed in the explants obtained from fully-opened flowers. High frequency shoot regeneration was also obtained from the petal explants of cvs. Coral. Lena, Nora and White Sim, and an interspecific cultivar Eolo using the method developed in this study.Abbreviations NAA
-naphthaleneacetic acid
- BA
6-benzyladenine
- GA3
gibberellic acid
- 2iP
N6-2-isopentenyl adenine
- KT-30
N-2-chloro-4-pyridyl-N-phenylurea (also called 4PU)
- TDZ
N-1,2,3-thiadiazol-5-yl-N-phenylurea (also called thidiazuron) 相似文献
11.
Summary The multiplet structure of cross peaks in double-quantum-filtered COSY NMR spectra is analysed for those resonances that include passive heteronuclear couplings. Interestingly, the cross peak involving the sugar-ring protons H2 and H3 in nucleic acids display an E.COSY-type appearance exclusively when the backbone torsion angle (C4-C3-O3-P) adopts a gauche(-) conformation. This observation allows an unambiguous analysis of the conformation around , without the knowledge of 3Jcp coupling constants. 相似文献
12.
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
13.
Summary Organic pyrophosphates such as UppA and NAD are formed when a solution containing a nucleotide, a nucleoside 5-polyphosphate, Mg2+ and imidazole are allowed to dry out. We suggest that this synthesis may have occured concurrently with oligonucleotide formation.Abbreviations Im
Imidazole
- CDI
1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride
- EDTA
ethylenediaminetetraacetic acid
- A
adenosine
- U
uridine
- pnA
adenosine 5-poly-phosphate containing n phosphate residues
- pU
uridine 5-phosphate
- AppA
P1,P2-diadenosine 5-pyrophosphate
- UppA
P1-(uridine 5)-P2-(adenosine 5)-pyrophosphate
- ImpA
adenosine 5-phosphorimidazolide
- NMN
nicotinamide mononucleotide
- NAD
nicotinamide-adenine dinucleotide 相似文献
14.
V. A. Wagner T. A. Schild H. Geldermann 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,89(1):121-126
For the detection of polymorphisms within the 5-flanking region of the -lactoglobulin (-LG) -encoding gene a nucleotide sequence containing 795 bp of the promoter and 59 bp of exon I was cloned and sequenced. After comparing the sequence from the DNA of 11 diverse cows (different breeds and milk-protein yields), 14 singlebp substitutions were identified within the 5-flanking region and two in the 5-untranslated region (5-UTR) of exon I. Some of the variants are located in potential binding sites for trans-acting factors or in the 5-UTR. A PCR-based RFLP analysis was performed, and the genotypes of an additional 60 cows were identified at five variable 5-flanking sites. The results reveal three frequent combinations between the A and B alleles of the protein-coding region and the novel 5-flanking DNA variants. This finding may explain the differences of the protein-variant-dependent -LG synthesis (A>B) observed in vivo. A sequence comparison of the bovine and ovine promoters reveals an homology of 92.8% and shows a higher degree of conservation between positions -600 and -300. 相似文献
15.
Ahlert Schmidt 《Archives of microbiology》1977,112(3):263-270
Crude extracts of Rhodospirillum rubrum catalyzed the formation of acid-volatile radioactivity from (35S) sulfate, (35S) adenosine-5-phosphosulfate, and (35S) 3-phosphoadenosine-5-phosphosulfate. An enzyme fraction similar to APS-sulfotransferases from plant sources was purified 228-fold from Rhodospirillum rubrum. It is suggested here that this enzyme is specific for adenosine-5-phosphosulfate, because the purified enzyme fraction metabolized adenosine-5-phosphosulfate, however, only at a rate of 1/10 of that with adenosine-5-phosphosulfate. Further, the reaction with 3-phosphoadenosine-5-phosphosulfate was inhibited with 3-phosphoadenosine-5-phosphate whereas this nucleotide had no effect on the reaction with adenosine-5-phosphosulfate. For this activity with adenosine-5-phosphosulfate the name APS-sulfotransferase is suggested. This APS-sulfotransferase needs thiols for activity; good rates were obtained with either dithioerythritol or reduced glutathione; other thiols like cysteine, 2-3-dimercaptopropanol or mercaptoethanol are less effective. The electron donor methylviologen did not catalyze this reaction. The pH-optimum was about 9.0; the apparent K
m for adenosine-5-phosphosulfate was determined to be 0.05 mM with this so far purified enzyme fraction. Enzyme activity was increased with K2SO4 and Na2SO4 and was inhibited by 5-AMP. These properties are similar to assimilatory APS-sulfotransferases from spinach and Chlorella.Abbreviations APS
adenosine-5-phosphosulfate
- PAPS
3-phosphoadenosine-5-phosphosulfate
- 5-AMP
adenosine-5-monophosphate
- 3-AMP
adenosine-3-monophosphate
- 3-5-ADP
3-phosphoadenosine-5-phosphate (PAP)
- DTE
dithiorythritol
- GSH
reduced glutathione
- BAL
2-3-dimercaptopropanol 相似文献
16.
Biosynthetic studies with cell-free extracts from Aloe arborescens Mill. demonstrate the transfer of the glucose moiety from UDP-glucose to aloe emodin anthrone, forming the C-glycosidic linkage in the anthracene derivative aloin. The pH-dependence and the specificity of UDP-glucose and aloe emodin anthrone for the biosynthesis of the C-glycosidic bond in aloin are shown.Abbreviations ADP-Glc
adenosine-5-diphosphate glucose
- AEA
aloe emodin anthrone (1,8-dihydroxy-3-(hydroxymethyl)-9(10 H)-anthracenone)
- CoASAc
acetyl coenzyme A
- GDP-Glc
guanosine-5-diphosphate glucose
- Glc
glucose
- Glc-1-P
glucose-1-phosphate
- HPLC
high performance liquid chromatography
- TLC
thin layer chromatography
- UDP-Gal
uridine-5-diphosphate galactose
- UDP-Glc
uridine-5-diphosphate glucose 相似文献
17.
Summary 2-Amino-2-deoxyuridine reacts efficiently with nucleoside 5-phosphorimidazolides in aqueous solution. The dinucleoside monophosphate analogues were obtained in yields exceeding 80% under conditions in which little reaction occurs with the natural nucleosides.In a similar way, the 5-phosphorimidazolide of 2-amino-2-deoxyuridine undergoes self-condensation in aqueous solution to give a complex mixture of oligomers.The phosphoramidate bond in the dinucleoside monophosphate analogues is stable for several days at room temperature and pH 7. The mechanisms of their hydrolysis under acidic and alkaline conditions are described.Abbreviations A
adenosine
- C
cytidine
- G
guanosine
- U
uridine
- T
thymidine
- UN
3
2-azido-2-deoxyuridine
- UNH
2
2-amino-2-deoxyuridine
- ImpA
adenosine 5-phosphorimidazolide
- ImpU
uridine 5-phosphorimidazolide
- ImpUN
3
2-azido-2-deoxyuridine 5-phosphorimidazolide
- ImpUNH
2
2-amino-2-deoxyuridine 5-phosphorimidazolide
- pA
adenosine 5-phosphate
- pU
uridine 5-phosphate
- pUN
3
2-azido-2-deoxyuridine 5-phosphate
- pUNH
2
2-amino-2-deoxyuridine 5-phosphate
- UpA
uridylyl-[35]-adenosine
- UpU
uridylyl-[35]-uridine
- UNpA
adenylyl-[52]-2-amino-2-deoxy-uridine
- UNpU
uridylyl-[52]-2-amino-2-deoxyuridine (pUN)n n=2,3,4 [25]-linked oligomers of pUNH
2 poly(A) polyadenylic acid
- Im
imidazole
- MeIm
l-methylimidazole 相似文献
18.
J. Visscher R. van der Woerd C. G. Bakker Alan W. Schwartz 《Origins of life and evolution of the biosphere》1989,19(1):3-6
Evidence is presented that a poly(U) template selectively favors the oligomerization of the activated, 3–5 pyrophosphate-linked dimer pdAppdAp, in comparison with the 3–3 and 5–5 linked dimers. In the absence of poly(U), the 5–5linked dimer is the most reactive, and chains are formed which are more than 60 monomer units in length.Nucleic Acid-Like Structures V. For the previous paper in this series see Visscher and Schwartz (1988). 相似文献
19.
Johannes H. Ippel Virginia Lanzotti Aldo Galeone Luciano Mayol Jacquelien E. van den Boogaart Jeroen A. Pikkemaat Cornelis Altona 《Journal of biomolecular NMR》1995,6(4):403-422
Summary The circular DNA decamer 5-dpCGC-TT-GCG-TT-3 was studied in solution by means of NMR spectroscopy and molecular dynamics in H2O. At a temperature of 269 K, a 50/50 mixture of two dumbbell structures (denoted L2L2 and L2L4) is present. The L2L2 form contains three Watson-Crick C-G base pairs and two two-residue loops in opposite parts of the molecule. On raising the temperature from 269 K to 314 K, the L2L4 conformer becomes increasingly dominant (95% at 314 K). This conformer has a partially disrupted G(anti)-C(syn) closing base pair in the 5-GTTC-3 loop with only one remaining (solvent-accessible) hydrogen bond between NH of the cytosine dC(1) and O6 of the guanine dG(8). The opposite 5-CTTG-3 loop remains stable. The two conformers occur in slow equilibrium (rate constant 2–20 s–1). Structure determination of the L2L2 and L2L4 forms was performed with the aid of a full relaxation matrix approach (IRMA) in combination with restrained MD. Torsional information was obtained from coupling constants. Coupling constant analysis (3JHH, 3JHP, 3JCP) gave detailed information about the local geometry around backbone torsion angles , , and , revealing a relatively high flexibility of the 5-GTTC-3 loop. The values of the coupling constants are virtually temperature-independent. Weakly constrained molecular dynamics in solvent was used to sample the conformational space of the dumbbell. The relaxation matrices from the MD simulation were averaged over r–3 to predict dynamic NOE volumes. In order to account for the 1:1 conformational mixture of L2L2 and L2L4 present at 271 K, we also included S2 factors and r–6 averaging of the r–3-averaged relaxation matrices. On matrix averaging, the agreement of NOE volumes with experiment improved significantly for protons located in the thermodynamically less stable 5-GTTC-3 loop. The difference in stability of the 5-CTTG-3 and 5-GTTC-3 loops is mainly caused by differences in the number of potential hydrogen bonds in the minor groove and differences in stacking overlap of the base pairs closing the minihairpin loops. The syn conformation for dC(1), favored at high temperature, is stabilized by solvation in the major groove. However, the conformational properties of the dC(1) base, as deduced from R-factor analysis and MD simulations, include a large flexibility about torsion angle . 相似文献
20.
D. S. Domozych 《Protoplasma》1989,149(2-3):108-119
Summary Cytochemical analysis of the endomembrane system of the chlamydomonad flagellate,Gloeomonas kupfferi (Chlorophyta), reveals distinct compartmentalization. Phosphatase localization shows that: IDPase is located throughout all cisternae of the dictyosome and vesicles associated with the contractile vacuole. Other alkaline phosphatases like TPPase, ATPase and ITPase were localized within the trans-face cisternae and vesicles of the contractile vacuole. IMPase was localized at the plasmamembrane and not within the endomembrane system. Acid phosphatases, incl. CMPase, NADPase and -glycerophosphatase, were localized in vesicles emerging from the central terminus of the trans-face of the dictyosome and in the peripheral vacuolar network. Silver proteinate labeling was noted in the dictyosome, contractile vacuole and on the anterior plasmamembrane. A summary of endomembrane compartmentalization and a putative interpretation of membrane flow and economy are presented.Abbreviations ER
endoplasmic reticulum
- IDPase
inosine 5-diphosphatase
- ITPase
inosine 5-triphosphatase
- ATPase
adenosine 5-triphosphatase
- TPPase
thiamine pyrophosphatase
- CMPase
cytidine 5-monophosphatase
- NADPase
-nicotinamide adenine diphosphatase
- AcPase
acid phosphatase 相似文献