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1.
Summary A method for measuring three-bond 13C-1H scalar coupling constants across glycosidic bonds in a cyclic (12)-glucan icosamer is presented. This oligosaccharide molecule, with its high degree of symmetry, represents a particular challenge for NMR spectroscopy to distinguish inter-residue from intra-residue heteronuclear coupling effects. Chemically equivalent H2 protons in adjacent glucosyl residues are distinguished on the basis of their different through-space, dipolar interactions with the anomeric protons (H1). The strong NOE contact between anomeric (H1) and aglyconic (H2) protons permits the selective observation of the inter-residue heteronuclear couplings 3JC1H2 and 3JC2H1 in a natural-abundance 13C-1-half-filtered {1H,1H} ROESY experiment.Abbreviations COSY
scalar correlated spectroscopy
- NOE
nuclear Overhauser effect
- NOESY
NOE spectroscopy
- ROESY
rotating-frame NOE spectroscopy 相似文献
2.
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
3.
Ariane van der Straten Rosette Loriau Albert Herzog Alex Bollen 《Bioscience reports》1986,6(4):363-373
Various constructions of human haptoglobin (Hp) cDNA coding either for the complete 2FS precursor protein or only for the subunit have been placed under the control of the PR promoter in the bacterial expression vector pCQV2 (Queen, 1983). In addition to the expected 45,000 dalton polypeptide synthesized after induction of the PR promoter, the complete 2FS constructions constitutively express a smaller polypeptide of 30,000 dalton corresponding to a truncated Hp protein. Computer analysis of the HpcDNA revealed the presence of two strong potential bacterial promoters (2PF and 2PS) located in the duplicated 2FS sequence. Both Hp promoter signals are followed by potential mRNA start sites and ribosome binding sites at a compatible distance from initiation codons. In addition, the Hp2 cDNA sequence, when fused upstream to the cDNA coding for 1-antitrypsin, constitutively promotesin vivo the efficient expression of an hybrid protein specifically recognized by antibodies raised against 1-antitrypsin or haptoglobin. 相似文献
4.
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
5.
C. Mark Fletcher David N. M. Jones Robert Diamond David Neuhaus 《Journal of biomolecular NMR》1996,8(3):292-310
Summary Two modifications to the commonly used protocols for calculating NMR structures are developed, relating to the treatment of NOE constraints involving groups of equivalent protons or nonstereoassigned diastereotopic protons. Firstly, a modified method is investigated for correcting for multiplicity, which is applicable whenever all NOE intensities are calibrated as a single set and categorised in broad intensity ranges. Secondly, a new set of values for pseudoatom corrections is proposed for use with calculations employing centre-averaging. The effect of these protocols on structure calculations is demonstrated using two proteins, one of which is well defined by the NOE data, the other less so. It is shown that failure to correct for multiplicity when using r-6 averaging results in overly precise structures, higher NOE energies and deviations from geometric ideality, while failure to correct for multiplicity when using r-6 summation can cause an avoidable degradation of precision if the NOE data are sparse. Conversely, when multiplicities are treated correctly, r-6 averaging, r-6 summation and centre averaging all give closely comparable results when the structure is well defined by the data. When the NOE data contain less information, r-6 averaging or r-6 summation offer a significant advantage over centre averaging, both in terms of precision and in terms of the proportion of calculations that converge on a consisten result.Abbreviations HMG
high mobility group
- NOE
nuclear Overhauser enhancement
- NOESY
nuclear Overhauser enhancement spectroscopy
- rmsd
root-mean-square deviation
- YASAP
yet another simulated-annealing protocol 相似文献
6.
Leszek Poppe Rainer Stuike-Prill Bernd Meyer Herman van Halbeek 《Journal of biomolecular NMR》1992,2(2):109-136
Summary We present a comprehensive strategy for detailed characterization of the solution conformations of oligosaccharides by NMR spectroscopy and force-field calculations. Our experimental strategy generates a number of interglycosidic spatial constraints that is sufficiently large to allow us to determine glycosidic linkage conformations with a precision heretofore unachievable. In addition to the commonly used {1H,1H} NOE contacts between aliphatic protons, our constraints are: (a) homonuclear NOEs of hydroxyl protons in H2O to other protons in the oligosaccharide, (b) heteronuclear {1H,13C} NOEs, (c) isotope effects of O1H/O2H hydroxyl groups on13C chemical shifts, and (d) long-range heteronuclear scalar coupling across glycosidic bonds.We have used this approach to study the trisaccharide sialyl-(26)-lactose in aqueous solution. The experimentally determined geometrical constraints were compared to results obtained from force-field calculations based on Metropolis Monte Carlo simulations. The molecule was found to exist in 2 families of conformers. The preferred conformations of the (26)-linkage of the trisaccharide are best described by an equilibrium of 2 conformers with angles at –60° or 180° and of the 3 staggered rotamers of the angle with a predominantgt conformer. Three intramolecular hydrogen bonds, involving the hydroxyl protons on C8 and C7 of the sialic acid residue and on C3 of the reducing-end glucose residue, contribute significantly to the conformational stability of the trisaccharide in aqueous solution.
Supplementary material available from the corresponding author: Table containing values for the dihedral angles , , , , and for bond angles , for the six lowest-energy conformations of sialyl-(26)-lactose (1 page). 相似文献
7.
The reaction of the 5 -phosphorimidazolide of adenosine (5-ImpA) with diadenosine pyrophosphate (A5ppA) in the presence of Na+-montmorillonite in aqueous, pH 8 solution results in the regiospecific formation of A5ppA3pA and A5ppA3pA3 pA. The formation of oligomers of general structure (pA)n decreases in the presence of A5ppA. A5ppA3pA is the principal reaction product when a 1:1 ratio of ImpA and A5ppA is used. The yield of A5ppA3pA3pA is optimal when 9:1 or 4:1 ratios of ImpA: A5ppA are used. The overall regiospecificity of formation of 3,5-links is about 80%. The reaction between ImpA and A5ppA on montmorillonite differs from the self-condensation of ImpA in that it proceeds in the absence of Mg2+ and there are only small differences in oligomer yields when Na+, Li+ Ca2+, and NH
4
+
are the exchangeable cations on the montmorillonite. The reaction is inhibited by 0.4 M imidazole but the inhibition is suppressed with 0.4 M Mg2+. Little or no phosphodiester bond formation was observed with Mg2+- or Al3+-montmorillonite. Montmorillonites other than 22A and Volclay exhibited no catalysis for the formation of adducts between ImpA and A5ppA and no catalysis was exhibited in ferrugenous smectite, nontronite, allophane, or sepiolite. 相似文献
8.
Jorge Santoro Marta Bruix Carlos González José L. Nieto Manuel Rico 《Journal of biomolecular NMR》1992,2(6):647-653
Summary A constant-time version of the homonuclear NOESY experiment (CT-NOESY) is described. The experiment yields simplified protein spectra, in which cross peaks arising from protons with zero or small couplings are differentiated from other cross peaks, thus partially overcoming the problem of signal overlap. In addition, the CT-NOESY spectrum provides information on the magnitude of3JNH- and3J coupling' constants, and is thus useful to determine torsion angle constraints and to perform stereospecific assignments of CHH protons in the case of3J constants. 相似文献
9.
Carlomagno T Maurer M Sattler M Schwendinger MG Glaser SJ Griesinger C 《Journal of biomolecular NMR》1996,8(2):161-170
Summary A new homonuclear Hartmann-Hahn-type mixing scheme is introduced that effects coherence transfer between resonances in two separated frequency bands. The mixing scheme relies on the irradiation of two-band selective shaped pulses that are expanded in an MLEV-16 supercycle. Similar to heteronuclear Hartmann-Hahn experiments, a planar effective coupling tensor is created. This novel mixing scheme is applied to C,C transfer and to the transfer between C and aromatic carbon spins. 相似文献
10.
G. S. Sidhu G. S. Khush 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1978,53(5):199-203
Summary The inheritance of resistance to brown planthopper, Nilaparvata lugens (Stol.), of 20 rice cultivars was studied. Single dominant genes that are allelic to Bph 3 condition the resistance in cultivars Ptb 19, Gangala (Acc. 7733), Gangala (Acc. 15207), Horana Mawee, Kuruhondarwala, Mudu Kiriyal and Muthumanikam. Single recessive genes that are allelic to bph 4 govern the resistance in cultivars Gambada Samba, Heenhoranamawee, Hotel Samba, Kahata Samba, Kalukuruwee, Lekam Samba, Senawee, Sulai, Thirissa and Vellai Illankali. The resistance in Ptb 33, Sudu Hondarwala, and Sinna Sivappu is governed by one dominant and one recessive gene which segregate independently of each other. The dominant resistance genes in these cultivars appear allelic to either Bph 1 or Bph 3. Similarly, the recessive genes in these cultivars seem allelic to either bph 2 or bph 4. Further investigations are needed to conclusively determine the allelic relationships of resistance genes in Ptb 33, Sudu Hondarwala and Sinna Sivappu. 相似文献
11.
Claudia P. Spampinato Piotr Paneth Marion H. O'Leary Carlos S. Andreo 《Photosynthesis research》1991,28(2):69-76
Structural analogues of the NADP+ were studied as potential coenzymes and inhibitors for NADP+ dependent malic enzyme from Zea mays L. leaves. Results showed that 1, N6-etheno-nicotinamide adenine dinucleotide phosphate ( NADP+), 3-acetylpyridine-adenine dinucleotide phosphate (APADP+), nicotinamide-hypoxanthine dinucleotide phosphate (NHDP+) and -nicotinamide adenine dinucleotide 2: 3-cyclic monophosphate (23NADPc+) act as alternate coenzymes for the enzyme and that there is little variation in the values of the Michaelis constants and only a threefold variation in Vmax for the five nucleotides. On the other hand, thionicotinamide-adenine dinucleotide phosphate (SNADP+), 3-aminopyridine-adenine dinucleotide phosphate (AADP+), adenosine 2-monophosphate (2AMP) and adenosine 2: 3-cyclic monophosphate (23AMPc) were competitive inhibitors with respect to NADP+, while -nicotinamide adenine dinucleotide 3-phosphate (3NADP+), NAD+, adenosine 3-monophosphate (3AMP), adenosine 2: 5-cyclic monophosphate (25AMPc), 5AMP, 5ADP, 5ATP and adenosine act as non-competitive inhibitors. These results, together with results of semiempirical self-consistent field-molecular orbitals calculations, suggest that the 2-phosphate group is crucial for the nucleotide binding to the enzyme, whereas the charge density on the C4 atom of the pyridine ring is the major factor that governs the coenzyme activity.Abbreviations NADP+
1, N6-etheno-nicotinamide adenine dinucleotide phosphate
- NHDP+
nicotinamide-hypoxanthine dinucleotide phosphate
- APADP+
3-acetylpyridine-adenine dinucleotide phosphate
- SNADP+
thionicotinamide-adenine dinucleotide phosphate
- AADP+
3-aminopyridine-adenine dinucleotide phosphate
- 23NADPc+
-nicotinamide adenine dinucleotide 2: 3-cyclic monophosphate
- 3NADP+
-nicotinamide adenine dinucleotide 3-phosphate
- 2AMP
adenosine 2-monophosphate
- 3AMP
adenosine 3-monophosphate
- 23AMPc
adenosine 2: 3 monophosphate cyclic
- A
adenosine
- RuBP
ribulose 1,5-bisphosphate
- SCF-MO
Self-Consistent Field-Molecular Orbitals (method) 相似文献
12.
Philip H. Johnson Alastair S. R. Donald Julia L. Clarke Winifred M. Watkins 《Glycoconjugate journal》1995,12(6):879-893
1,3-Fucosyltransferase solubilized from human liver has been purified 40 000-fold to apparent homogeneity by a multistage process involving cation exchange chromatography on CM-Sephadex, hydrophobic interaction chromatography on Phenyl Sepharose, affinity chromatography on GDP-hexanolamine Sepharose and HPLC gel exclusion chromatography. The final step gave a major protein peak that co-chromatographed with 1,3-fucosyltransferase activity and had a specific activity of 5–6 µmol min–1 mg–1 and anM
r 44 000 deduced from SDS-PAGE and HPLC analysis. The purified enzyme readily utilized Gal1-4GlcNAc, NeuAc2-3Gal1-4GlcNAc and Fuc1-2Gal1-4GlcNAc, with a preference for sialylated and fucosylated Type 2 acceptors. Fuc1-2Gal1-4Glc and the Type 1 compound Gal1-3GlcNAc were very poor acceptors and no incorporation was observed with NeuAc2-6Gal1-4GlcNAc. A polyclonal antibody raised against the liver preparation reacted with the homologous enzyme and also with the blood group Lewis gene-associated 1,3/1,4-fucosyltransferase purified from the human A431 epidermoid carcinoma cell line. No cross reactivity was found with 1,3-fucosyltransferase(s) isolated from myeloid cells. Examination by Northern blot analysis of mRNA from normal liver and from the HepG2 cell line, together with a comparison of the specificity pattern of the purified enzyme with that reported for the enzyme expressed in mammalian cells transfected with theFuc-TVI cDNA, suggests a provisional identification ofFuc-TVI as the major 1,3-fucosyltransferase gene expressed in human liver.Died June, 1991 相似文献
13.
Dong-Hyun Kim Norihiro Azuma Hideyuki Tanaka Choemon Kanno 《Glycoconjugate journal》1998,15(4):361-369
The structures of the N-linked sugar chains in the PAS-6 glycoprotein (PAS-6) from the bovine milk fat globule membrane were determined. The sugar chains were liberated from PAS-6 by hydrazinolysis, and the pyridylaminated sugar chains were separated into a neutral (6N) and two acidic chains (6M and 6D), the acidic sugar chains then being converted to neutral sugar chains (6MN and 6DN). 6N was separated into two neutral fractions (6N13 and 6N5.5), while 6MN and 6DN each gave a single fraction (6MN13 and 6DN13). The structure of 6N5.5, which was the major sugar chain in PAS-6, is proposed to be Man16 (Man13) Man14GlcNAc14GlcNAc-PA; 6N13, 6MN13 and 6DN13 are proposed to be Gal13Gal14GlcNAc12Man16 (Gal13Gal14GlcNAc12Man13) Man14GlcNAc14 (Fuc16)GlcNAc-PA;6M and 6D had 1 or 2 additional NeuAc residues at the non-reducing ends of 6MN13 and 6DN13, respectively. © 1998 Rapid Science Ltd 相似文献
14.
P. R. K. Koduru M. K. Rao 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1980,56(3):137-143
Summary Thirteen spontaneously occurring chlorophyll deficient phenotypes have been described and their genetic basis was established. Ten of these — white, white tipped green, patchy white, white virescent, white striping 1, white striping 2, white striping 4, fine striping, chlorina and yellow virescent showed monogenic recessive inheritance and the remaining three — yellow striping, yellow green and light green seedling phenotypes showed digenic recessive inheritance. The genes for (i) white tipped green (wr) and yellow virescent (yv) and (ii) patchy white (pw) and white striping 1 (wst 1) showed independent assortment. Further, the genes for white (w), white tipped green (wr) and yellow virescent (yv) were inherited independently of the gene for hairy leaf margin (Hm).In the mutants — white tipped green, patchy white, white striping 1, white striping 2, fine striping, chlorina, yellow virescent, yellow striping, yellow green and light green phenotypes total quantity of chlorophyll was significantly less than that in the corresponding controls, while in white virescent there was no reduction in the mature stage. For nine of the mutants the quantity of chlorophyll was also estimated in F1's (mutant x control green). In F1's of six of the mutants — white tip, patchy white, chlorina, yellow virescent, fine striping and yellow striping the quantity of chlorophyll was almost equal to the wild type. In the F1's of three of the mutants — white striping 1, white striping 2 and light green an intermediate value between the mutant and wild types was observed. In yellow virescent retarded synthesis of chlorophyll, particularly chlorophyll a was observed in the juvenile stage. Reduced quantity of chlorophyll was associated with defective chloroplasts. In the mutants — white tipped green, white virescent, fine striping, chlorina, yellow striping, yellow green and light green defective plastids were also observed. In patchy white secondary destruction of chlorophylls and the presence of defective plastids were found to be associated with reduced chlorophyll quantity at maturity.Paper chromatographic studies of leaf flavonoids revealed some variation between the inbreds, but there were three common spots, 7, 8 and 9, except for PDP in which the spot 8 was absent. Chlorophyll deficient mutants differed from their respective controls in the absence of one or more of the spots present in the controls and in the presence of new spots in some of the mutants.Most of the chlorophyll mutants showed higher survival rate in the Kharif season than in Rabi season which was attributed to the higher mean day temperature and longer day light period in the Kharif season than in Rabi season. 相似文献
15.
Bellizzi Mary C. Dutta-Roy Asim K. James W. Philip T. 《Molecular and cellular biochemistry》1997,170(1-2):187-193
The role of -tocopherol uptake system in human erythrocyte in the uptake of plasma -tocopherol has been suggested. However no information is available on -tocopherol uptake activity of human erythrocytes in the presence of high levels of D-glucose which is known to lead to pathological alterations in different cells including human erythrocytes. Therefore, in order to examine the effect of D-glucose on the binding of -tocopherol to human erythrocytes, the binding characteristics of -tocopherol to these cells were established first. Binding of [3H]-tocopherol to human erythrocytes was both saturable and specific. Scatchard analysis of -tocopherol binding to these cells showed the presence of two independent classes of binding sites with widely different affinities. The high affinity binding sites had a dissociation constant (Kd1) of 90 nM with a binding capacity (n1) of 900 sites per cell, whereas the low affinity binding sites had a dissociation constant (Kd2) of 5.2 M and a binding capacity (n2) of 105,400 sites per cell. Trypsin treatment abolished all the -tocopherol binding activity. Competition for the binding of -tocopherol to human erythrocytes was effective with other homologues of -tocopherol (-tocopherol, -tocopherol and -tocopherol) and their potency was almost equal to -tocopherol itself. The order of preference was -tocopherol > -tocopherol -tocopherol -tocopherol. Incubation of human erythrocytes with various concentrations of D-glucose did not affect -tocopherol uptake activity. Our data demonstrate the presence of an -tocopherol uptake system in human erythrocytes and that the -tocopherol uptake activity is not modulated by the presence of D-glucose. 相似文献
16.
N. N. Smirnov 《Hydrobiologia》1966,27(3-4):337-343
Summary The differences in the structure of the trunk limbs allow to outline three sections of Chydoridae (see table I and fig. 1), coinciding with the sections distinguished according to the structure of the head pores.
Chydoridae (Cladocera)
Chydoridae (. ), , .相似文献
17.
Summary Long-range physical maps of the small multigene family of the malt -amylase genes (-Amy-1) located on the long arms of wheat chromosomes 6A (the -Amy-A1 locus) and 6B (-Amy-B1) were generated by pulsed-field gel electrophoresis analysis. By using three methylation-sensitive rare-cutter restriction endonucleases, NotI, NruI and MluI, and an -Amy-1 cDNA probe and four gene-specific genomic probes from the -Amy-B1 locus, the size of the -Amy-B1 locus was estimated to be about 700 kb and of the -Amy-B1 locus to be about approximately 4300 kb. These two maps indicate clustering of GC-rich and C-methylation-sensitive restriction enzyme recognition sites. At least five regions reminiscent of CpG islands are apparent in -Amy-B1, and three in -Amy-A1. Correlation between recombination frequency and physical distance within the -Amy-B1 locus suggests that 1 cM approximates to 1 Mb in physical distance. 相似文献
18.
Summary A frequency-time domain 3D NMR technique has been developed for measurement of heteronuclear coupling constants in oligonucleotides employing a combination of COSY andJ-resolved techniques. The method employs frequency-selective excitation to generate the 1 axis and 2D FT to generate the 2 and 3 axes. The procedure yields high resolution, especially along the 1 axis. The technique is demonstrated on a dinucleotide. 相似文献
19.
Summary Isolated amphibian (Triturus alpestris) gastrula ectoderm was treated with cyclic nucleotides for 24 h and cultured up to 12 days. Explants treated with$cyclic N6-Monobutyryl-adenosine-35-monophosphate, cyclic Dibutyryladenosine-35-monophosphate and cyclic Dibutyrylguanosine-35-monophosphate in a concentration of 10–3 and 10–5 M did not differentiate into mesoderm- or endoderm-derived tissues. The number of explants with small neural and neuroid structures did not exceed the percentage found in the control series. Inductions could also not be obtained when ectoderm was dissociated prior to the treatment with cyclic nucleotides, or when theophylline (which inhibits phosphodiesterase) was added to the culture medium. The results are discussed with regard to the possible mode of action of the vegetalizing factor. 相似文献
20.
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