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1.
Tadashi II Masayuki Kubota Satoshi Okuda Takashi Hirano Mamoru Ohashi 《Glycoconjugate journal》1995,12(2):162-172
Negative-ion fast atom bombardment tandem mass spectrometry has been used in the characterization of non-, mono-, di- and trisulfated disaccharides from heparin and heparan sulfate. The positional isomers of the sulfate group of monosulfated disaccharides were distinguished from each other by negative-ion fast atom bombardment tandem mass spectra, which provide an easy way of identifying the positional isomers. This fast atom bombardment collision induced dissociation mass spectrometry/mass spectrometry technique was also applied successfully to the characterization of di- and trisulfated disaccharides.Abbreviations FABMS
fast atom bombardment mass spectrometry
- CID
collision induced dissociation
- MIKE
mass analysed ion kinetic energy
- MS/MS
mass spectrometry/mass spectrometry
- HPLC
high performance liquid chromatography
- UA
d-gluco-4-enepyranosyluronic acid
- CS
chondroitin sulfate
- DS
dermatan sulfate
- HA
hyaluronan
- Hep
heparin
- HS
heparan sulfate
- UA(14) GlcNAc
2-acetamido-2-deoxy-4-O-(-d-gluco-4-enepyranosyluronic acid)-d-glucose
- UA(14)GlcNAc6S
2-acetamido-2-deoxy-4-O-(-d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-glucose
- UA2S(14)GlcNAc
2-acetamido-2-deoxy-4-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-d-glucose
- UA2S(14)GlcNAc6S
2-acetamido-2-deoxy-4-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-glucose
- UA(14)GlcN6S
2-amino-2-deoxy-4-O-(-d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-glucose
- UA2S(14)GlcN
2-amino-2-deoxy-4-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-d-glucose
- UA2S(14)GlcN6S
2-amino-2-deoxy-4-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-glucose
- UA(14)GlcNS
2-deoxy-2-sulfamino-4-O-(-d-gluco-4-enepyranosyluronic acid)-d-glucose
- UA(14)GlcNS6S
2-deoxy-2-sulfamino-4-O-(-d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-glucose
- UA2S(14)GlcNS
2-deoxy-2-sulfamino-4-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-d-glucose
- UA2S(14)GlcNS6S
2-deoxy-2-sulfamino-4-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-glucose
- UA(13)GalNAc
2-acetamido-2-deoxy-3-O-(-d-Gluco-4-enepyranosyluronic acid)-d-galatose
- UA(13)GalNAc4S
2-acetamido-2-deoxy-3-O-(-d-gluco-4-enepyranosyluronic acid)-4-O-sulfo-d-galactose
- UA(13)GalNAc6S
2-acetamido-2-deoxy-3-O-(-d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-galactose
- UA2S(13)GalNAc
2-acetamido-2-deoxy-3-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-d-galactose
- UA2S(13)GalNAc4S
2-acetamido-2-deoxy-3-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-4-O-sulfo-d-galactose
- UA2S(13)GalNAc6S
2-acetamido-2-deoxy-3-O-(2-O-sulfo--d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-galactose
- UA(13)GalNAcDiS
2-acetamido-2-deoxy-3-O-(-d-gluco-4-enepyranosyluronic acid)-4,6-di-O-sulfo-d-galactose
- UA(13)GlcNAc
2-acetamido-2-deoxy-3-O-(-d-gluco-4-enepyranosyluronic acid)-d-glucose. 相似文献
2.
P. N. Marshall 《The Histochemical journal》1976,8(5):487-499
Synopsis Commercial samples of Erythrosin B (CI 45430), Erythrosin Y (CI 45425), Fluorescein (CI 45350), Phloxine (CI 45410) and Rose Bengal (CI 45440) have been analysed by thin-layer chromatography. The Erythrosins were found to be mixtures consisting in the main of 4-iodofluorescein, 4,5-di-iodofluorescein, 2,4,5-triiodofluorescein and 2,4,5,7-tetraiodofluorescein, in some instances together with 2,4,5-tri-iodo-4,5,6,7-tetrachlorofluorescein and 2,4,5,7-tetraiodo-4,5,6,7-tetrachlorofluorescein. Samples of Fluorescein were mixtures of the nominal dye usually with traces of several unidentified, fluorescent components. Those of Phloxine consisted mainly of mixtures of 4-bromo-4,5,6,7-tetrachlorofluorescein, 4,5-dibromo-4,5,6,7-tetrachlorofluorescein, 2,4,5-tribromo-4,5,6,7-tetrachlorofluorescein and 2,4,5,7-tetrabromo-4,5,6,7-tetrachlorofluorescein, often with 4,5,6,7-tetrachlorofluorescein Samples of Rose Bengal were mixtures of 4-iodo-4,5,6,7-tetrachlorofluorescein, 4,5-di-iodo-4,5,6,7-tetrachlorofluorescein, 2,4,5-tri-iodo-4,5,6,7-tetrachlorofluorescein and 2,4,5,7-tetraiodo-4,5,6,7-tetrachlorofluorescein together with some unidentified components.Most of the commercial dye samples gave an insoluble residue when extracted with methanol. This residue was usually inorganic carbonate or halide. Some possible practical consequences of the various impurities are discussed. 相似文献
3.
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
4.
Karl Hård Albert Mekking Johannis P. Kamerling Georges A. A. Dacremont Johannes F. G. Vliegenthart 《Glycoconjugate journal》1991,8(1):17-28
Five brain-derived and 17 urinary oligomannose-type oligosaccharides were isolated by ion-exchange chromatography on Mono Q or Dowex, followed by HPLC on Lichrosorb-NH2 from a Persian cat suffering from -mannosidosis. The structures ofthe carbohydrate chains were determined by 500- or 600-MHz1H-NMR spectroscopy. Different oligosaccharide patterns were found in brain and urine. 99% of the urinary oligosaccharides possess an (1-6)-linked mannose residue attached to -mannose, whereas only 5% of the brain-derived oligosaccharides contain such a residue. Furthermore, of the urinary carbohydrate chains 71% end with Man1-4GlcNAc1-4GlcNAc and 29% end with Man1-4GlcNAc, whereas the corresponding amounts are 23% and 77%, respectively, for the brain-derived oligosaccharides.Abbreviations MLEV-17
composite pulse devised by M. Levitt
- HOHAHA
homonuclear Hartman-Hahn spectroscopy
- TPPI
time-proportional phase incrementation
- 2D
two dimensional
- GlcNAc
N-acetylglucosamine
- Man
mannose
- Fuc
fucose 相似文献
5.
Fumito Matsuura Masaya Ohta Khoichi Murakami Yujirou Matsuki 《Glycoconjugate journal》1993,10(3):202-213
Structures of the Asn linked oligosaccharides of quail egg-yolk immunoglobulin (IgY) were determined in this study. Asn linked oligosaccharides were cleaved from IgY by hydrazinolysis and labelled withp-aminobenzoic acid ethyl ester (ABEE) afterN-acetylation. The ABEE labelled oligosaccharides were then fractionated by a combination of Concanavalin A-agarose column chromatography and anion exchange, normal phase and reversed phase HPLC before their structures were determined by sequential exoglycosidase digestion, methylation analysis, HPLC, and 500 MHz1H-NMR spectroscopy. Quail IgY contained only neutral oligosaccharides of the following categories: the glucosylated oligomannose type (0.6%, Glc1-3Glc1-3Man9GlcNAc2; 35.6%, Glc1-3Man7–9GlcNAc2). oligomannose type (15.0%, with the structure Man5–9GlcNAc2) and biantennary complex type with core structures of-Man1-3(-Man1-6)Man1-4GlcNAc1-4GlcNAc (9.9%),-Man1-3(GlcNAc1-4)(-Man1-6)Man1-4GlcNAc1-4GlcNAc (25.1%) and-Man1-3(GlcNAc1-4)(-Man1-6)Man1-4GlcNAc1-4(Fuc1-6)GlcNAc (11.4%). Although never found in mammalian proteins, glucosylated oligosaccharides (Glc1Man7–9GlcNAc2) have been located previously in hen IgY.Abbreviations IgG, IgM, IgA, IgY
immunoglobulin G, M, A and Y, respectively
- ABEE
p-aminobenzoic acid ethyl ester 相似文献
6.
Rudolf Geyer Silvia Diabaté Hildegard Geyer Hans-Dieter Klenk Heiner Niemann Stephan Stirm 《Glycoconjugate journal》1987,4(1):17-32
Fowl plague virus, strain Dutch, was metabolically labeled withd-[2-3H]mannose, or withd-[6-3H]glucosamine, and the small subunit (HA2; 0.8 mg in total) of the viral hemagglutinin was isolated by preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis. After proteolytic digestion, the radioactive oligosaccharides were sequentially liberated from the glycopeptides by treatment with different endo--N-acetylglucosaminidases and with peptide:N-glycosidase or, finally, by hydrazinolysis. In this manner, four groups of glycans could be obtained by consecutive gel filtrations and were subfractionated by HPLC. The structures of the individual oligosaccharides were analyzed by micromethylation, by acetolysis or by digestion with exoglycosidases. The major species amongst the high mannose glycans at Ans-406 of the viral glycopolypeptide were found to be Man1-2Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNac1-4GlcNAc and Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNAc1-4GlcNAc, while the complex glycans at Asn-478 are predominantly GlcNAc1-2Man1-3(GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc (lacking, in part, one of the outerN-acetylglucosamine residues) and GlcNAc1-2Man1-3(Gal1-4GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc.Abbreviation BSA
bovine serum albumin
- endo D (F,H)
endo--N-acetyl-d-glucosaminidase D (F,H)
- HA
hemagglutinin (HA1, large subunit of HA
- HA2
small subunit
- FPV
fowl plague virus
- PNGase F
peptide:N-glycosidase F
- SDS
sodium dodecylsulfate 相似文献
7.
Eneyskaya EV Kulminskaya AA Kalkkinen N Nifantiev NE Arbatskii NP Saenko AI Chepurnaya OV Arutyunyan AV Shabalin KA Neustroev KN 《Glycoconjugate journal》2001,18(10):827-834
An -L-fucosidase (E.C. 3.2.1.51) exhibiting a wide aglycon specificity expressed in ability of cleaving 1 6-, 1 3-, 1 4-, and 1 2-O-fucosyl bonds in fucosylated oligosaccharides, has been isolated from culture filtrate of Thermus sp. strain Y5. The -L-fucosidase hydrolyzes p-nitrophenyl -L-fucopyranoside with V
max of 12.0 ± 0.1 M/min/mg and K
m = 0.20 ± 0.05 mM and is able to cleave off about 90% of total L-fucose from pronase-treated fractions of fucosyl-containing glycoproteins and about 30% from the native glycoproteins. The purified enzyme is a tetramer with a molecular mass of 240 ± 10 kDa consisting of four identical subunits with a molecular mass of 61.0 ± 0.5 kDa. The N-terminal sequence showed homology to some -L-fucosidases from microbial and plant sources. Hydrolysis of p-nitrophenyl -L-fucopyranoside occurs with retention of the anomeric configuration. Transglycosylating activity of the -L-fucosidase was demonstrated in reactions with such acceptors as alcohols, N-acetylglucosamine and N-acetylgalactosamine while no transglycosylation products were observed in the reaction with p-nitrophenyl -L-fucopyranoside. The enzyme can be classified in glycosyl hydrolase family 29. 相似文献
8.
Fumiaki Nakatsubo T. Kent Kirk Mikio Shimada Takayoshi Higuchi 《Archives of microbiology》1981,128(4):416-420
The degradation of the phenylcoumaran substructure model compound methyl dehydrodiconiferyl alcohol by the white-rot wood decay fungus Phanerochaete chrysosporium was investigated using culture conditions optimized for lignin oxidation. Initial attack was in the cinnamyl alcohol side chain, which was oxidized to a glycerol structure. This was subsequently converted by loss of the two terminal carbon atoms, C and C, to yield a C-aldehyde structure, which was further oxidized to the C-acid compound. The next detected intermediate, a phenylcoumarone, was produced by double bond formation between C and C, and oxidation of the C-alcohol to an aldehyde group. Further oxidation of C to an acid yielded the next intermediate. The final identified degradation product was veratric acid. No products from the 5-substituted aromatic ring, and no phenolic products, were found. The initial glycerol-containing intermediate was a mixture of the threo and erythro forms, and no optical activity could be found, suggesting that its formation might have involved nonstereospecific C-C epoxidation followed by non-enzymatic hydrolysis of the epoxide.Abbreviations TLC
thin layer chromatography
- LDA
lithium diisopropyl amide
- DDQ
2,3-dichloro-5,6-dicyanobenzoquinone
- MS
mass spectrometry
- UV
ultraviolet spectroscopy 相似文献
9.
The specific features of the topology of spectral powers and coherent interregional interrelationships in the narrow, individually determined -, -, 1-, 2-, and 3-frequency bands were studied by means of high-resolution EEG (62 channels) in novice and experienced meditators (NMs and EMs) at rest and under the conditions of generation of an altered state of consciousness characterized by inactivation of cognitive activity and the occurrence of a positive emotional experience of happiness. EMs in the meditation-free state were found to be characterized by a shift in the values of the individual frequency to a lower-frequency region of the spectrum, along with higher, compared to NMs, -, 1-, 2-, and 3-band power values, which probably reflects the cumulative character of the influence of long-term meditative practice. The effective achievement of altered states of consciousness in EMs was associated with an increase in the local - and 1 powers in the anterior cortical areas, as well as long-distance coherence between the prefrontal and posterior associative cortex with the formation of a center of gravity in the left prefrontal region (lead AF
3). According to the data of the correlation analysis of the EEG power values and the data of subjective scaling of the meditation state, the -power values were positively associated with positive emotional experiences and negatively associated with the level of mental activity. The results of this study are consistent with current concepts that the and activities in narrow frequency bands reflect the activity of multifunctional neuronal networks selectively associated with processes of cognitive and affective activity. 相似文献
10.
Klaus Steube Volker Gross Wolfgang Hösel Thuy-anh Tran-Thi Karl Decker Peter C Heinrich 《Glycoconjugate journal》1986,3(3):247-254
Endo--N-acetylglucosaminidase F (endo F, EC 3.2.1.96) and peptide:N-glycosidase F (PNGase F, EC 3.2.2.18) fromFlavobacterium meningosepticum were used for the deglycosylation of 1-proteinase inhibitor and 1-acid glycoprotein carrying oligosaccharide side chains of the complex-, high-mannose- and hybrid-type. High-mannose-and hybrid-type glycoproteins were obtained by the incubation of rat hepatocyte primary cultures with 1-deoxymannojirimycin or swainsonine, respectively. It was found that endo F cleaves hybrid- and high-mannose-type 1-proteinase inhibitor and 1-acid glycoprotein at pH 4.5 as well as at pH 8.5 in the presence or absence of 1% octyl--d-glucopyranoside. Complex-type 1-proteinase inhibitor or 1-acid glycoprotein were not cleaved by endo F even in the presence of octyl--d-glucopyranoside.PNGase F was found to cleave complex-, hybrid- and high-mannose-type oligosaccharide side chains of 1-proteinase inhibitor and 1-acid glycoprotein at pH 4.5 and pH 8.5 in the presence of 0.75% octyl--d-glucopyranoside. The deglycosylation of both protein substrates was very poor without detergents.Abbreviations Endo F
endo--N-acetylglucosaminidase F (EC 3.2.1.96)
- PNGase F
peptide:N-glycosidase F (EC 3.2.2.18)
Dedicated to Prof. Dr. Wolfgang Gerok on the occasion of his 60th birthday 相似文献
11.
G. P. Vlasov V. I. Korol'kov G. A. Pankova I. I. Tarasenko A. N. Baranov P. B. Glazkov A. V. Kiselev O. V. Ostapenko E. A. Lesina V. S. Baranov 《Russian Journal of Bioorganic Chemistry》2004,30(1):12-20
We attempted to find some compounds for the effective delivery of gene constructs into cells and obtained two trispherical dendrimers on the basis of lysine, (Lys)8-(,-Lys)4-(,-Lys)2-(,-Lys)-Ala-NH2 (D1) and ((Lys)8-(,-Lys)4-(,-Lys)2-,-Lys)-Ala-[Lys(Plm)]2-Ala-NH2 (D2), as well as the starburst polymeric derivatives of D1, (pVIm)
8
-D1 and (pLys)
n
-D1, containing poly(N-vinylimidazole) and polylysine chains single-point bound to the dendrimer amino groups. The conditions of dendrimer–plasmid DNA complex formation were studied. The intracellular localization of these complexes and the expression of gene constructs delivered with their help were analyzed in transfection experiments on the HeLa cell cultures of human epithelial carcinoma and on mouse C2C12 myoblasts. It was found that the chemical structure of dendrimer D1 and its derivatives significantly affected the structure and properties of complex. 相似文献
12.
J. Desmet E. Tieghem H. Van Dael E. Van Cauwelaert 《European biophysics journal : EBJ》1991,20(5):263-268
By means of reaction calorimetry we measured the apparent enthalpy change, Happ, of the binding of Mn2+-ions to goat -lactalbumin as a function of temperature. The observed Happ can be written as the sum of contributions resulting from a conformational and a binding process. In combination with the thermal unfolding curve of goat -lactalbumin, we succeeded in separating the complete set of thermodynamic parameters (H, G, S, Cp) into the binding and conformational contributions. By circular dichroism we showed that NH
4
+
-ions, upon binding to bovine a-lactalbumin, induce the same conformational change as do Na+ and K+: the binding constant
equals 98 ± 9 M–1.Abbreviations BLA
bovine -lactalbumin
- GLA
goat -lactalbumin
- HLA
human -lactalbumin
- CD
circular dichroism
Offprint requests to: H. Van DaelDeceased 相似文献
13.
Gijsbertus W. van Eijk Henri J. Roeijmans Hubertus A. van der Aa 《Antonie van Leeuwenhoek》1988,54(4):325-330
Clumps of white crystals present in 40-day-old malt agar cultures of Holwaya mucida were isolated as long white needles by crystallization from ethanol following short extraction with chloroform. The levorotary compound ([]
289
21
=-193.8°) was recognized as a -lactone (C17H20O5) by infrared and mass spectrometry. It was identified as 7-methoxy-3a, 10b-dimethyl-1, 2, 3, 3a, 5a, 7, 10b, 10c-octahydro-4H, 9H-furo[2, 3, 4 : 4, 5]naphtho[2, 1-c]pyran-4, 9-dione, a labdane-derived compound known as antibiotic LL-Z1271. Preparative thin-layer chromatography of the mother liquor afforded 2 minor metabolites. One was identified as LL-Z1271, the demethylated analogue of LL-Z1271. The other one named LL-Z1271, was recognized as a compound related to and : its structure could not be fully elucidated.
H. mucida (anamorph: Crinula calciiformis) has no taxonomic relationship with two other LL-Z1271 producing species viz. Acrostalagmus sp. (= Acremonium cf. atrogriseum) and Oidiodendron truncatum. 相似文献
14.
Summary Soluble lead salts and a number of lead-containing minerals catalyze the formation of oligonucleotides from nucleoside 5-phosphorimidazolides. The effectiveness of lead compounds correlates strongly with their solubility. Under optimal conditions we were able to obtain 18% of pentamer and higher oligomers from ImpA. Reactions involving ImpU gave smaller yields.Abbreviations A
adenosine
-
U
uridine
-
Im
imidazole
-
MeIm
1-methyl-imidazole
-
EDTA
ethylenediaminetetraacetic acid
-
pA
adenosine 5-phosphate
-
pU
uridine 5-phosphate
-
Ap
adenosine cyclic 2:3-phosphate
-
ATP
adenosine 5-triphosphate
-
AppA
P1,P2-diadenosine 5-diphosphate
-
pNp (N = A,U)
nucleotide 2(3), 5-diphosphate
-
ImpA
adenosine 5-phosphoreimidazolide
-
ImpU
uridine 5-phosphorimidazolide
-
A
2pA
adenylyl-[25]-adenosine
-
A
3pA
adenylyl-[35]-adenosine
-
pA
2pA
5-phospho-adenylyl-[25]-adenosine
-
pA
3pA
5-phospho-adenylyl-[35]-adenosine
-
pUpU
5-phospho-uridylyl-uridine
-
pApU
5-phospho-adenylyl-uridine
-
pUpA
5-phospho-uridylyladenine
-
(pA)n (n, 2,3,4,)
oligoadenylates with 5 terminal phosphate
-
ImpApA
5-phosphorimidazolide of adenylyl adenosine
-
(pA)
5+
pentamer and higher oligoadenylates with 5 terminal phosphate
-
(Ap)nA (n = 2,3,4)
oligoadenylates without terminal phosphates
In the following we do not specify the nature of the internucleotide linkageIn the following we do not specify the nature of the internucleotide linkage 相似文献
15.
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
16.
Summary The purified bacteriophage replication proteins O and P sediment separately in metrizamide gradients of low ionic strength as dimers. Together they interact with each other forming an oligomer, composed of two molecules of O and one molecule of P. The O-P oligomer is active in the in vitro replication of ori-containing DNA.Equilibrium sedimentation in preformed metrizamide density gradients under conditions that separate DNA-protein complexes from free proteins was employed in order to study possible interactions among the replication proteins and ori DNA. It was found that the P protein binds specifically to ori-containing plasmid DNA only in the presence of O protein. About 100 molecules of O and 10 molecules of P form a complex with the ori DNA. The DNA-O-P complex was shown to be active in an in vitro replication system.Since the physical interactions between ori and O and between P and the Escherichia coli dnaB replication protein are well documented, the evidence for a O-P interaction presented in this paper provides the missing link in the molecular mechanism that enables to direct the host replication machinery to the replication of its own DNA. 相似文献
17.
Debrabant Alain Bastien Patrick Dwyer Dennis M. 《Molecular and cellular biochemistry》2001,220(1-2):109-116
Previously, we isolated and characterized the gene encoding the 3-Nucleotidase/Nuclease (Ld3NT/NU) from the human pathogen, Leishmania donovani. This unique cell surface enzyme has been shown to be involved in the salvage of host-derived purines, which are essential for the survival of this important protozoan parasite. In this report, we assessed whether the 3-Nucleotidase/Nuclease was conserved amongst other pathogenic Leishmania and related trypanosomatid parasites. Results of pulsed field gel electrophoresis and Southern blotting showed that a Ld3NT/NU gene homolog was present in each of the visceral and cutaneous Leishmania species tested (i.e. isolates of L. donovani, L. infantum, L. tropica, L. major and L. mexicana, respectively). Further, results of colorimetric assays using 3-adenosine monophosphate as substrate demonstrated that each of these organisms also expressed significant levels of 3-nucleotidase enzyme activity. In addition, we showed that a Ld3NT/NU gene homolog was expressed in each of these Leishmania species as a > 40 kDa 3-nucleotidase enzyme activity. A Ld3NT/NU gene homolog was also identified in two Crithidia species (C. fasciculata and C. luciliae) and Leptomonas seymouri but was only marginally detectable in Trypanosoma brucei, Trypanosoma cruzi and Phytomonas serpens. Cumulatively, results of this study showed that an Ld3NT/NU homolog was conserved amongst pathogenic Leishmania sp. which suggests that this enzyme must play an critical role in purine salvage for all members of this group of human pathogens. 相似文献
18.
Takenori Miyamoto Diego Restrepo Edward J. Cragoe Jr. John H. Teeter 《The Journal of membrane biology》1992,127(3):173-183
Summary Olfactory receptor neurons enzymatically dissociated from channel catfish olfactory epithelium were depolarized transiently following dialysis of IP3 or cAMP (added to the patch pipette) into the cytoplasm. Voltage and current responses to IP3 were blocked by ruthenium red, a blocker of an IP3-gated Ca2+-release channel in sarcoplasmic reticulum. In contrast, the responses to cAMP were not blocked by extracellularly applied ruthenium red, nor by l-cis-diltiazem or amiloride and two of its derivatives. The current elicited by cytoplasmic IP3 in neurons under voltage clamp displayed a voltage dependence different from that of the cAMP response which showed marked outward rectification. A sustained depolarization was caused by increased cytoplasmic IP3 or cAMP when the buffering capacity for Ca2+ of the pipette solution was increased, when extracellular Ca2+ was removed or after addition of 20–200 nm charibdotoxin to the bathing solution, indicating that the repolarization was caused by an increase in [Ca
i
] that opened Ca2+-activated K+ channels. The results suggest that different conductances modulated by either IP3 or cAMP are involved in mediating olfactory transduction in catfish olfactory receptor neurons and that Ca2+-activated K+ channels contribute to the termination of the IP3 and cAMP responses.Abbreviations ATP
adenosine 5-triphosphate
- BAPTA
(bis-(o-aminophenoxy)-ethane-N-N-N-N)-tetraacetic acid
- cAMP
adenosine cyclic 3,5-monophosphate
- cGMP
guanosine cyclic 3,5-monophosphate
- CTX
charybdotoxin
- DCB
3,4-dichlorobenzamil
- EDTA
ethylenediaminetetraacetic acid
- EGTA
ethylenglycol-bis-(b-aminoethyl)-N-N-N-N-tetraacetic acid
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- IP3
inositol-1,4,5-triphosphate
- NMDG
N-methyl-d-glucamine
We would like to thank the Tanabe Seiyaku Co., Ltd., for their gift of l-cis-diltiazem. This work was supported by National Institutes of Health grants DC00566 and BRSG S07RR05825. 相似文献
19.
Sa-nga Pootrakul Suwat Sapprapa Prawase Wasi Supa Na-Nakorn Romsai Suwanik 《Human genetics》1975,29(2):121-126
Summary In the Far East two types of -thalassemia genes, namely -thalassemia1 (-thal1) and -thalassemia2 (-thal2) exist. Definite diagnosis of the -thal1 and -thal2 traits is very difficult because their hematological findings are minimally abnormal or normal. This study attempts to characterize the heterozygotes by hemoglobin chain synthesis in reticulocytes from obligatory cases of the -thal1 and -thal2 traits. Twelve parents of babies with hemoglobin Bart's hydrops fetalis (obligatory -thal1 trait) had the mean total radioactivity / ratio of 0.76±SD 0.04, while that of 7 normal controls was 1.06±SD 0.04. The / globin chain ratios of 16 cases, who were either parents or offspring of patients with hemoglobin H disease, were found to segregate into 2 groups, i.e. 0.78±SD 0.03 (10 cases) and 0.92±SD 0.03 (6 cases), probably representing the -thal1 and -thal2 traits respectively. The hematological data of the first group showed definite hypochromic microcytic red cells, similar to thoseof the parents of the hydrops. The second group had significantly higher mean corpuscular hemoglobin than the first group, compatible with -thal2 trait. Our globin chain synthesis study thus appears to be capable of discriminating normal, -thal1 and -thal2 traits.A preliminary report of the results was presented at the XV Congress of the International Society of Haematology, Israel, September, 1974. 相似文献
20.
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