共查询到20条相似文献,搜索用时 171 毫秒
1.
Irene M.A. Nooren Ke-Yu Wang Philip N. Borer István Pelczer 《Journal of biomolecular NMR》1998,11(3):319-328
The subject RNA models the binding site for the coat protein of the R17 virus, as well as the ribosome recognition sequence for the R17 replicase gene. With an RNA of this size, overlaps among the sugar protons complicate assignments of the 1H NMR spectrum. The cross peaks that overlap significantly in 2D-NOE spectra can frequently be resolved by introducing a third, in our approach the double-quantum, frequency axis. In particular the planes in a 3D-NOE/2QC spectrum perpendicular to the 2Q axis are extremely useful, showing a highly informative repeating NOE-2Q pattern. In this experiment substantial J-coupling confers special advantages. This always occurs for geminal pairs (H5/H5 for RNA plus H2/H2 for DNA), as well as for H5/H6, for H3/H4 in sugars with substantial populations of the N-pucker, for H1/H2 for S-puckered sugars, and usually for H2/H3. For the 24-mer RNA hairpin the additional information from the 3D-NOE/2QC spectrum allowed assignment of all of the non-exchangeable protons, eliminating the need for stable-isotope labeling. 相似文献
2.
Shcheglovitov A. K. Zhelay T. I. Kondratskii A. P. Naidenov V. G. Shuba Ya. M. 《Neurophysiology》2004,36(2):93-101
We analyzed the effects of nifedipine on a family of recombinant low-threshold Ca2+ channels functionally expressed in Xenopus oocytes and formed by three different subunits (1G, 1H, and 1I). The 1G and 1I channels demonstrated a low sensitivity to nifedipine even in high concentrations (IC50 = 98 and 243 M, maximum blocking intensity Amax = 25 and 47%, respectively). At the same time, the above agent effectively blocked channels formed by the 1H-subunit (IC50 = 5 M and Amax = 41%). The nifedipine-caused effects were voltage-dependent, and their changes depended on the initial state of the channel. In the case of 1G-subunits, the blockade was determined mostly by binding of nifedipine with closed channels, whereas in the cases of 1H- and 1I-subunits this resulted from binding of nifedipine with channels in the activated and inactivated states. The obtained data allow us to obtain estimates of the pharmacological properties of the above three subtypes of recombinant channels and, in the future, to compare these characteristics with the properties of low-threshold Ca2+ channels in native cells. 相似文献
3.
Cecilia PC Soh Alastair SR Donald James Feeney Walter TJ Morgan Winifred M Watkins 《Glycoconjugate journal》1989,6(3):319-332
The tetrasaccharides GalNAcß1-4[NeuAc2-3]Galß1-4Glc and GalNAcß1-4[NeuAc2-3]Galß1-4GlcNAc were synthesised by enzymic transfer of GalNAc from UDP-GalNAc to 3-sialyllactose (NeuAc2-3Galß1-4Glc) and 3-sialyl-N-acetyllactosamine (NeuAc2-3Galß1-4GlcNAc). The structures of the products were established by methylation and1H-500 MHz NMR spectroscopy. In Sda serological tests the product formed with 3-sialyl-N-acetyllactosamine was highly active whereas that formed with 3-sialyllactose had only weak activity. 相似文献
4.
Internodal cells of the giant alga Chara corallina were perfused internally to replace the native cytoplasm, tonoplast and vacuole with artificial cytoplasm. Sodium efflux from perfused cells, measured by including 22Na in the perfusion media, was increased by increasing the internal sodium concentration and by decreasing the external pH, and was inhibited by external application of the renal diuretic amiloride. The sodium efflux was markedly ATP-dependent, with a 50-fold decrease in efflux observed after perfusion with media lacking ATP. Efflux in the presence of ATP was reduced by 33% by inclusion of 10 M N,N-dicyclohexylcarbodiimide in the perfusion medium. The membrane potential of the perfused cells approximated that of intact cells from the same culture. It is suggested that sodium efflux in perfused Chara cells proceeds via a secondary antiporter with protons, regulated by ATP in a catalytic role and with the proton motive force acting as the energy source.Abbreviations DCCD
N,N-dicyclohexylcarbodiimide
- EGTA
ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid
- Mes
2-(N-morpholino)ethanesulphonic acid
- Mops
3(N-morpholino)propanesulphonic acid
- Taps
tris(hydroxymethyl)methylaminopropanesulphonic acid 相似文献
5.
M. Dutta V. Arunachalam A. Bandyopadhyay K. Vinod Prabhu 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1986,71(4):662-666
Summary Seeds of 4 crosses of groundnut (Arachis hypogaea L.), Robut 33-1 x Chico, Robut 33-1 x NC Ac 17090, Robut 33-1 x PI 298115 and MK 374 x GAUG 1, were irradiated with 30 kR. In the F1, some branches of each plant were intermated with other plants at random and others selfed in each cross to produce S2 and F2 seeds. They were evaluated for pod yield, shelling percentage and 100-kernel weight. The frequency of plants superior to F[in1] was much higher in S2 than in F2, which was, in general, true for the values of yield and its components. The S2 and F2 were advanced to third generation by selfing. The families descending from S2 showed clear superiority over those from F2. The reason for such superiority was suggested to be the recombination of genes from the upper and lower ends of the genotypic distribution under intermating. 相似文献
6.
Six tetrasaccharide fractions were isolated from shark cartilage chondroitin sulfate D by gel filtration chromatography followed by HPLC on an amine-bound silica column after exhaustive digestion with testicular hyaluronidase. Their structures were determined unambiguously by one- and two-dimensional 500 MHz1H NMR spectroscopy in conjunction with HPLC analysis of chondroitinase AC-II digests of the tetrasaccharides. One fraction was found to contain two tetrasaccharide components. All the seven tetrasaccharides shared the common core structure GlcA1-3GalNAc1-4GlcA1-3GalNAc with various sulfation profiles. Four were disulfated comprising of two monosulfated disaccharide units GlcA1-3GalNAc(4-sulfate) and/or GlcA1-3GalNAc(6-sulfate), whereas the other three were hitherto unreported trisulfated tetrasaccharides containing a disulfated disaccharide unit GlcA(2-sulfate)1-3GalNAc(6-sulfate) and a monosulfated disaccharide unit GlcA1-3GalNAc(4-or 6-sulfate). These sulfated tetrasaccharides were demonstrated to serve as appropriate acceptor substrates for serum -N-acetylgalactosaminyltransferase, indicating their usefulness as authentic oligosaccharide substrates or probes for the glycobiology of sulfated glycosaminoglycans.Abbreviations NFU
National formulary unit
- COSY
correlation spectroscopy
- HOHAHA
homonuclear Hartmann-Hahn
- 1D or 2D
one- or two-dimensional
- IdoA
l-iduronic acid
- GlcA
d-gluco-4-enepyranosyluronic acid
- Di-0S
GlcA1-3GalNAc
- Di-4S
GlcA1-3GalNAc(4-sulfate)
- Di-4S
GlcA1-3GalNAc(4-sulfate)
- Di-6S
GlcA1-3GalNAc(6-sulfate)
- Di-6S
GlcA1-3GalNAc(6-sulfate)
- Di-diS
d
GlcA(2-sulfate)1-3GalNAc(6-sulfate)
- Di-diSE
GlcA1-3GalNAc(4, 6-disulfate)
- U
G, U, 2S, 4S, and 6S represent GlcA, GalNAc, GlcA, 2-O-sulfate, 4-O-sulfate, and 6-O-sulfate, respectively 相似文献
7.
Kenwyn R. Gayler Sotirios Kolivas Alison J. Macfarlane Glenn G. Lilley Mauro Baldi Robert J. Blagrove Elizabeth D. Johnson 《Plant molecular biology》1990,15(6):879-893
The biosynthesis of conglutin has been studied in developing cotyledons of Lupinus angustifolius L. Precursors of conglutin formed the major sink for [35S]-cysteine incorporated by developing lupin cotyledons, and these precursors were rapidly sequestered into the endoplasmic reticulum. The sequence of a cDNA clone coding for one such precursor of conglutin was determined. The structure of the precursor polypeptide for conglutin predicted from the cDNA sequence contained an N-terminal leader peptide of 22 amino acids directly preceding a subunit polypeptide of M
r 4520, together with a linking region of 13 amino acids and a subunit polypeptide of M
r 9558 at the C-terminus. The amino acid sequence predicted from the cDNA sequence showed minor variations from that established by sequencing of the protein purified from mature dried seeds (Lilley and Inglis, 1986). These were consistent with the existence of a multi-gene family coding for conglutin . Comparison of the sequences of conglutin with those of other 2S storage proteins showed that the cysteines involved in internal disulphide bridges between the mature subunits of conglutin , were maintained throughout this family of proteins but that little else was conserved either at the protein or DNA level. 相似文献
8.
Gerard Strecker Jean-Michel Wieruszeski Jean-Claude Michalski Jean Montreuil 《Glycoconjugate journal》1988,5(4):385-396
The structure of a new nonasaccharide isolated from human milk has been investigated. By using methylation analysis, FAB-MS and1H-and13C-NMR spectroscopy as basic methods of structural investigation, this oligosaccharide was identified as VI2--Fuc,V4-Fuc,III3--Fuc-p-lacto-n-hexaose: Fuc1-2Gal1-3[Fuc1-4]GlcNAc1-3Gal1-4[Fuc1-3]GlcNAc1-3Gal1-4Glc.Abbreviations COSY
correlation spectroscope
- DP
degree of polymerisation
- FAB-MS
fast atom bombardment-mass spectrometry
- HPLC
high performance liquid chromatography
- NMR
nuclear magnetic resonance
- GLC
gas-liquid chromatography 相似文献
9.
Dr. R. V. Nair T. M. Masajo G. S. Khush 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1982,61(1):19-22
Summary The inheritance of resistance to whitebacked planthopper Sogatella furcifera (Horvath) was studied in 21 rice varieties. Reactions of F1; F2 and F3 progenies of the crosses of 21 resistant varieties with the susceptible variety TN 1 revealed that a single dominant gene governs resistance in Mushkan 41, Santhi, Siahnakidar 195, SM2-34, Tirisurkh 251, Zirijowaian 245, 18, 24A, 39, 76 S, 78, 180, 213 B, 267, 293, CI 6037-4, NP97, S39 JKW and Bansphul. In varieties 65 and 274 A, resistance is governed by one dominant and one recessive gene which segregate independently of each other. Tests for allelism with the Wbph 1 gene originally identified in N 22 revealed that the dominant gene present in all the test varieties is the same as Wbph 1. Further studies are required to determine the allelic relationships of the recessive gene found in varieties 65 and 274 A. 相似文献
10.
Unravelling the factors that contribute to the formation and the stability of -sheet structure in peptides is a subject of great current interest. A -hairpin, the smallest -sheet motif, consists of two antiparallel hydrogen-bonded -strands linked by a loop region. We have performed a statistical analysis on protein -hairpins showing that the most abundant types of -hairpins, 2:2, 3:5 and 4:4, have characteristic patterns of 13C and 13C conformational shifts, as expected on the basis of their and angles. This fact strongly supports the potential value of 13C and 13C conformational shifts as a means to identify -hairpin motifs in peptides. Their usefulness was confirmed by analysing the patterns of 13C and 13C conformational shifts in 13 short peptides, 10–15 residues long, that adopt -hairpin structures in aqueous solution. Furthermore, we have investigated their potential as a method to quantify -hairpin populations in peptides. 相似文献
11.
ATP synthase (F0F1) is driven by an electrochemical potential of H+ (H+). F0F1 is composed of an ion-conducting portion (F0) and a catalytic portion (F1). The subunit composition of F1 is 33. The active 33 oligomer, characterized by X-ray crystallography, has been obtained only from thermnophilic F1 (TF1). We proposed in 1984 that ATP is released from the catalytic site (C site) by a conformational change induced by the DELSEED sequence via -F0. In fact, cross-linking of DELSEED to stopped the ATP-driven rotation of in the center of 33. The torque of the rotation is estimated to be 420 pN·å from the H+ and H+-current through F0F1. The angular velocity () of is the rate-limiting step, because H+ increased theV
max of H+ current through F0, but not theK
m
(ATP). The rotational unit of F0 (=ab2c10) is /5, while that in 33 is 2/3. This difference is overcome by an analog-digital conversion via elasticity around DELSEED with a threshold to release ATP. The distance at the C site is about 9.6 å (2,8-diN3-ATP), and tight Mg-ATP binding in 33 was shown by ESR. The rotational relaxation of TF1 is too rapid (=100 nsec), but the rate of AT(D)P-induced conformational change of 33 measured with a synchrotron is close to . The ATP bound between the P-loop and E188 is released by the shift of DELSEED from RGL. Considering the viscosity resistance and inertia of the free rotor (-c), there may be a stator containing OSCP (= of TF1) and F0-d to hold free rotation of 33. 相似文献
12.
The transmembrane proton gradient of the sulfate-reducing bacterium Desulfovibrio desulfuricans strain CSN has been determined by in vivo31P nuclear magnetic resonance (NMR) spectroscopy in the absence of dioxygen. At pH 7.0 in the medium (pHex) the intracellular pH (pHin) was 7.5. By lowering pHex to 5.9 pHin decreased to 7.1. At pHex greater than 7.7 the transmembrane proton gradient (pH) was zero. The uncouplers 3,3,4,5-tetrachlorosalicylanilide (TCS) and carbonylcyanide-m-chlorophenylhydrazone (CCCP), or the permeant anion thiocyanate caused complete dissipation of pH.Abbreviations
CCCP
carbonylcyanide-m-chlorophenylhydrazone
-
TCS
3,3,4,5-tetrachlorosalicylanilide
-
MOPS
3-(N-morpholino)-propanesulfonic acid
-
P
i
inorganic phosphate
-
pH
in (pHex)
intracellular (extracellular) pH
- pH
transmembrane proton gradient (pHin-pHex)
-
electrochemical membrane potential
-
chemical shift in parts per million
-
NMR
nuclear magnetic resonance 相似文献
13.
Jennifer C. Wilson David C. M. Kong Yu-Teh Li Mark Von Itzstein 《Glycoconjugate journal》1996,13(6):927-931
The mechanism of hydrolysis of 4-methylumbelliferyl 3-deoxy-d-glycero--d-galacto-2-nonulopyranosidonic acid (KDN2MeUmb,4) by KDN-sialidase isolated from the hepatopancreas of the oysterCrassostrea virginica has been monitored by1H NMR spectroscopy. The results of these experiments reveal that KDN-sialidase catalyses the hydrolysis of the synthetic substrate KDN2MeUmb, with initial release of -d-KDN. This is consistent with an overall mechanism for the hydrolysis which proceeds with retention of anomeric configuration. These results agree with earlier NMR studies of otherN-acetylneuraminic acid-recognising sialidases from both viral and bacterial sources. 相似文献
14.
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
15.
Myobacterium avium LM1 was exposed to concentrations of 5-fluorouracil (5FU) that ranged from 0 to 100 g/ml. Growth inhibition was inversely proportional to the concentration of the drug. DNA was extracted from cells grown in medium that contained [14C]5FU, but no carrier. The [14C]DNA was enzymatically hydrolyzed to deoxyribonucleotides, which were separated and fractionated by high-performance liquid chromatography (HPLC). The isotope was located in 2-deoxycytidine 5-monophosphate (dCMP) and 2-deoxythymidine 5-monophosphate (dTMP), with dCMP containing the majority. There was no radioactivity at the elution times for 5-fluoro-2-deoxyuridine 5-monophosphate or 2-deoxyuridine 5-monophosphate. These results suggested that 5FU was dehalogenated and the uracil moiety ultimately converted into cytosine and thymine deoxyribonucleotides. Cells were grown in [3H]uracil, and [3H]DNA was extracted and analyzed by HPLC. The isotope was found only in the pyrimidine deoxyribonucleotides, with dCMP containing 4.1 times that in dTMP. Thus, it was demonstrated that uracil and dehalogenated 5FU were not directly incorporated into DNA, but rather converted to cytosine and thymine and then incorporated into DNA by a salvage pathway. 相似文献
16.
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
17.
Liping Yu Robert Goldman Patrick Sullivan Greg F. Walker Stephen W. Fesik 《Journal of biomolecular NMR》1993,3(4):429-441
Summary The structures of uniformly 13C-labeled -glucan octa- and undeca-oligosaccharides enzymatically prepared by the yeast cell wall glucanosyl transferase of Candida albicans were characterized by using a combination of HCCH-COSY, HCCH-TOCSY, and HMBC experiments. The oligosaccharide structures indicate that the cell wall glucanosyl transferase cleaves two glucosyl units from the reducing end of the initial linear (13) penta-oligosaccharide and subsequently transfers the remainder to another oligosaccharide at the nonreducing end via a (16) linkage. These results indicate that the combined action of cell wall glucanase and glucanosyl transferase activities could not only introduce intrachain (16) linkages within a single glucan strand, but also result in cross-linking of two initially separate glucan strands with concurrent introduction of intrachain (16) linkages. Since isolated fungal membranes only synthesize linear (13) glucan strands, wall-associated enzymes probably participate in the assembly of the final wall glucan structure during cell growth and division. 相似文献
18.
Christina Scha¨ffer Norbert Mu¨ller Pravat K. Mandal Rudolf Christian Sonja Zayni Paul Messner 《Glycoconjugate journal》2000,17(10):681-690
The peptidoglycan, the secondary cell wall polymer (SCWP), and the surface layer (S-layer) glycoprotein are the major glycosylated cell wall components of Paenibacillus alvei
CCM 2051. In this report, the complete structure of the SCWP, its linkage to the peptidoglycan layer, and its physicochemical properties have been investigated. From the combined evidence of chemical and structural analyses together with one- and two-dimensional nuclear magnetic resonance spectroscopy, the following structure of the SCWP-peptidoglycan complex is proposed:[(Pyr4,6)--D-Manp
NAc-(14)--D-Glcp
NAc-(13)]ñ11-(Pyr4,6)--D-Manp
NAc-(14)--D-Glcp
NAc-(1O)-PO2-O-PO2-(O6)-MurNAc-Each disaccharide unit is substituted by 4,6-linked pyruvic acid residues. Under mild acidic conditions, up to 50% of them are lost, leaving non-substituted ManNAc residues. The anionic glycan chains constituting the SCWP are randomly linked via pyrophosphate groups to C-6 of muramic acid residues of the peptidoglycan layer. 31P NMR reveals two signals that, as a consequence of micelle formation, experience different line broadening. Therefore, their integral ratio deviates significantly from 1:1. By treatment with ethylenediaminetetraacetic acid, sodium dodecyl sulfate, and sonication immediately prior to NMR measurement, this ratio approaches unity. The reversibility of this behavior corroborates the presence of a pyrophosphate linker in this SCWP-peptidoglycan complex.In addition to the determination of the structure and linkage of the SCWP, a possible scenario for its biological function is discussed. 相似文献
19.
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
20.
Elke Duchardt Christian Richter Bernd Reif Steffen J. Glaser Joachim W. Engels Christian Griesinger Harald Schwalbe 《Journal of biomolecular NMR》2001,21(2):117-126
A new heteronuclear NMR pulse sequence for the measurement of nJ(C,H) coupling constants, the /selective HC(C)H-TOCSY, is described. It is shown that the S3E element (Meissner et al., 1997a,b) can be used to obtain spin state selective coherence transfer in molecules, in which adjacent CH moieties are labeled with 13C. Application of the / selective HC(C)H-TOCSY to a 10nt RNA tetraloop 5-CGCUUUUGCG-3, in which the four uridine residues are 13C labeled in the sugar moiety, allowed measurement of two bond and three bond J(C,H) coupling constants, which provide additional restraints to characterize the sugar ring conformation of RNA in cases of conformational averaging. 相似文献