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1.
The white rot basidiomycete Phanerochaete chrysosporium metabolized 4-ethoxy-3-methoxyphenyl-glycerol--guaiacyl ether (V) in low nitrogen, stationary cultures under which conditions the ligninolytic enzyme system is expressed. 4-Ethoxy-3-methoxyphenylglycerol XIII, guaicol and 4-ethoxy-3-methoxybenzyl alcohol (II) were isolated as metabolic products. Exogenously added XIII was rapidly converted to 4-ethoxy-3-methoxybenzyl alcohol indicating that it is an intermediate in the metabolism of V. P. chrysosporium also metabolized 1-(4-ethoxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-3-hydroxypropane VI. The degradation pathway for this dimer also included initial -ether cleavage and -hydroxylation of the diol product 1-(4-ethoxy-3-methoxyphenyl) 2,3 dihydroxypropane (XI) to yield the triol XIII which was cleaved at the , bond to yield 4-ethoxy-3-methoxybenzyl alcohol. Finally P. chrysosporium also cleaved the dimer 1-(4-ethoxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-1-hydroxypropane (VIII) at the -ether linkage yielding 1-(4-ethoxy-3-methoxyphenyl) 1,2 dihydroxypropane (IX) which was subsequently cleaved at the , bond to yield II. All of the results indicate that oxidative -ether cleavage is an important initial reaction in the metabolism of -aryl ether lignin substructure dimeric compounds. Metabolities were identified after comparison with chemically synthesized standards by gas liquid chromatography-mass spectrometry.Abbreviations GLC
Gas liquid chromatography
- TMSi
trimethylsilyl
- TLC
thin layer chromatography 相似文献
2.
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
3.
Mycolic acid patterns of some species of Mycobacterium 总被引:5,自引:0,他引:5
David E. Minnikin S. Megan Minnikin James H. Parlett Michael Goodfellow Mogens Magnusson 《Archives of microbiology》1984,139(2-3):225-231
Representative strains of some species of Mycobacterium were degraded by both acid and alkaline methanolysis. Two-dimensional thin-layer chromatography was used to determine the patterns of mycolic acids and other long-chain components in these methanolysates. Patterns composed of -, methoxy- and ketomycolates were found in Mycobacterium asiaticum, Mycobacterium bovix, Mycobacterium gastri, Mycobacterium gordonae, Mycobacterium kansasii, Mycobacterium marinum and Mycobacterium tuberculosis; a representative of Mycobacterium thermoresistibile also contained lower molecular weight -mycolates in addition to these three acids. In representatives or Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium nonchromogenicum, Mycobacterium novum, Mycobacterium paratuberculosis, Mycobacterium scrofulaceum, Mycobacterium terrae, Mycobacterium xenopi, and Mycobacterium sp. MNC 165 - and ketomycolates were accompanied by -carboxymycolates and 2-eicosanol and homologous alcohols which are derived from wax-ester mycolates. Mycobacterium fortuitum and Mycobacterium giae contained - and epoxymycolates and both serovars of Mycobacterium simiae had a very characteristic pattern of -, - and ketomycolic acids. Comparison with data for other mycobacteria showed the chemotaxonomic significance of these mycolic acid patterns.Abbreviations TLC
thin-layer chromatography
- TBDMS
t-butyldimethylsilyl
Supplementary data: Copies of the chromatographic patterns of the mycolic acids from all the strains examined can be provided on request from one of the authors (D.E.M.) 相似文献
4.
Ben A. Bulthuis Gregory M. Koningstein Adriaan H. Stouthamer Henk W. van Verseveld 《Antonie van Leeuwenhoek》1993,63(1):1-16
The magnitude of the proton motive force (p) and its constituents, the electrical () and chemical potential (-ZpH), were established for chemostat cultures of a protease-producing, relaxed (rel
–) variant and a not protease-producing, stringent (rel
+) variant of an industrial strain ofBacillus licheniformis (respectively referred to as the A- and the B-type). For both types, an inverse relation of p with the specific growth rate was found. The calculated intracellular pH (pHin) was not constant but inversely related to . This change in pHin might be related to regulatory functions of metabolism but a regulatory role for pHin itself could not be envisaged. Measurement of the adenylate energy charge (EC) showed a direct relation with for glucose-limited chemostat cultures; in nitrogen-limited chemostat cultures, the EC showed an approximately constant value at low and an increased value at higher . For both limitations, the ATP/ADP ratio was directly related to .The phosphorylation potential (G'p) was invariant with . From the values for G'p and p, a variable H+/ATP-stoichiometry was inferred: H+/ATP=1.83+0.52µ, so that at a given H+/O-ratio of four (4), the apparent P/O-ratio (inferred from regression analysis) showed a decline of 2.16 to 1.87 for =0 to max (we discuss how more than half of this decline will be independent of any change in internal cell-volume). We propose that the constancy of G'p and the decrease in the efficiency of energy-conservation (P/O-value) with increasing are a way in which the cells try to cope with an apparent less than perfect coordination between anabolism and catabolism to keep up the highest possible with a minimum loss of growth-efficiency. Protease production in nitrogen-limited cultures as compared to glucose-limited cultures, and the difference between the A- and B-type, could not be explained by a different energy-status of the cells.Abbreviations CCCP
carbonylcyanide-p-trichloromethoxyphenylhydrazone
- DW
dry weight of biomass
- F
Faraday's constant, 96.6 J/(mV × mol)
- Fo
chemostat outflow-rate (ml/h)
- FCCP
carbonylcyanide-p-trifluoromethoxyphenylhydrazone
- G'p
phosphorylation potential, the Gibbs energy change for ATP-synthesis from ADP and Pi
- G'0p
standard Gibbs energy change at specified conditions
- H+/ATP
number of protons translocated through
- ATP
synthase in synthesis of one ATP
- H+/O
protons translocated during transfer of 2 electrons from substrate to oxygen
-
specific growth rate (1/h)
- H+
transmembrane electrochemical proton potential, J/mol
- Mb
molar weight (147.6 g/mol) of bacteria with general cell formula C6.0H10.8O3.0N1.2
- pHout,in
extracellular, intracellular pH
- Pi
(intracellular) inorganic phosphate
- p
proton motive force, mV
- pH
transmembrane pH-difference
-
transmembrane electrical potential, mV
- P/O
number of ADP phosphorylated to ATP upon reduction of one O2– to H2O by two electrons transferred through the electron transfer chain
- P/O
(H+/O) × (H+/ATP)–1
- P/OF, P/ON
P/O with the two electrons donated by resp. (NADH + H+) and FADH
- q
specific rate of consumption or production (mol/g DW × h)
-
rel
+,rel
–
stringent, relaxed genotype
- R
universal gas constant, 8.36 J/(mol × degree)
- T
absolute temperature
- TPMP+
triphenylmethylphosphonium ion
- TPP+
tetraphenyl phosphonium ion
- Y
growth yield, g DW/mol
- Z
conversion constant=61.8 mV for 310 K (37 °C)
- ZpH
transmembrane proton potential or chemical potential, mV 相似文献
5.
R. Garza C. Cardona S. P. Singh 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,92(3-4):357-362
The bean-pod weevil (BPW), Apion godmani Wagner, often causes heavy losses in crops of common bean (Phaseolus vulgaris L.). Farmers need resistant bean cultivars to minimize losses, cut production costs, stabilize seed yield, and reduce pesticide use and consequent health hazards. To design effective breeding methods, breeders need new and better sources of resistance and increased knowledge of their modes of inheritance. We therefore: (1) compared sources of resistance to BPW, (2) studied the inheritance of resistance, and (3) determined whether the sources possess similar or different genes for BPW resistance. The following sources of resistance, originating from the Mexican highlands, were evaluated for 3 years at INIFAP-Santa Lucía de Prias, Texcoco, Mexico: Amarillo 153, Amarillo 169, Hidalgo 58, J 117, Pinto Texcoco, Pinto 168, and Puebla 36. All except Puebla 36 were crossed with the susceptible cultivar Jamapa. Amarillo 153 and Puebla 36 were crossed with another susceptible cultivar, Bayo Mex. The parents, F1 hybrids, and F2 populations were evaluated for BPW damage in 1992. Backcrosses of the F1 of Jamapa/Pinto 168 to the respective susceptible and resistant parents were also evaluated in 1992. All seven resistant accessions were crossed in all possible combinations, excluding reciprocals. The resulting 21 F1 hybrids and 21 F2 populations were evaluated for BPW damage in 1994. J 117 had the highest level of resistance to BPW. Pinto Texcoco and Puebla 36 had the highest mean damage score of all seven sources of resistance. The F1 hybrids between susceptible parents and resistant sources were generally intermediate. Two genes segregating independently controlled the BPW resistance in each accession. One gene, Agm, has no effect when present alone, whereas the other gene, Agr, alone conferred intermediate resistance. When both genes were present, resistance to BPW was higher. Based on mean BPW damage scores, all 21 F1 hybrids and their F2 populations, derived from crosses among seven resistant accessions, were resistant. However, data from individual plant damage scores in F2 populations of Amarillo 169/Pinto 168 and Pinto Texcoco/Pinto 168 suggested that at least one gene in each of the three accessions was non-allelic. Data also indicated that Amarillo 169 had a dominant gene that conferred high levels of BPW resistance, irrespective of the alleles at the other locus; and that Pinto Texcoco and Pinto 168 possessed two different genes for intermediate resistance. 相似文献
6.
Nuclear and cytoplasmic gene control of resistance to loose smut (Ustilago tritici (Pers.) Rostr.) in wheat (Triticum aestivum L.) 总被引:1,自引:0,他引:1
P. Dhitaphichit P. Jones E. M. Keane 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1989,78(6):897-903
Summary Using disomic chromosome substitution lines based on the susceptible wheat cultivar Chinese Spring, loose smut resistance of wheat cultivars Hope and Thatcher was shown to be conferred in each case by a single dominant major gene carried on chromosome 7 A (Hope) or 7 B (Thatcher). Partial resistance was determined by genes on an additional eight Hope or seven Thatcher chromosomes, and similarities were evident between the partial resistance genotypes ofHope and Thatcher. Chinese Spring exhibited a mean infection value of approximately 50%, indicating a significant level of partial resistance, which was found to be due, in part, to genes on the homoeologous chromosome arms 1 As, 1 Es and 1 Ds, and to cytoplasmic genes. Substitution of the Chinese Spring nucleus into the cytoplasm of Aegilops squarrosa, Ae. variabilis or Ae. mutica resulted in increased susceptibility to Ustilago tritici. Several alloplasmic lines of the resistant wheat cultivars Selkirk and Chris exhibited race-specific susceptibility to U. tritici. 相似文献
7.
Our recent studies have revealed the existence of two distinct Gal: 3-O-sulfotransferases capable of acting on the C-3 position of galactose in a Core 2 branched structure, e.g., Gal14GlcNAc16(Gal13)GalNac1OBenzyl as acceptor to give 3-O-sulfoGal14GlcNAc13(Gal13)GalNAc1OB 20 and Gal14GlcNAc16(3-O-sulfoGal13)GalNAc1OB 23. We herein report the synthesis of these two compounds and also that of other modified analogs that are highly specific acceptors for the two sulfotransferases. Appropriately protected 1-thio-glycosides 7, 8, and 10 were employed as glycosyl donors for the synthesis of our target compounds. 相似文献
8.
M. Robert M. S. Noel-Hudson J. Font M. Aubery J. Wepierre 《Cell biology and toxicology》1994,10(5-6):361-365
Culture of keratinocytes on a noncoated porous synthetic membrane maintained at the air-liquid interface allows the establishment of a fibroblast/keratinocyte co-culture, without direct cell-cell contant between the two cellular layers. The influence of fibroblasts (proliferating, confluent or blocked by mitomycin C) on epidermization (i.e., expression of integrins and markers of epidermal differentiation) was studied by immunohistochemistry in two culture media. In the medium supplemented with VCS or Ultroser G and in the absence of fibroblasts, 2, 3, 5 and 6 subunits of integrins are expressed by the basal keratinocytes, except 5 which does not appear with the medium supplemented with Ultroser G. During stratification, the 3 subunit is the only one to persist on suprabasal cells and all the markers of epidermal differentiation studied (filaggrin, involucrin, transglutaminase, keratins K1/K10) are expressed at the 14th day of emerged culture. The presence of fibroblasts modifies the expression profile of integrins: when they are proliferative, the expression of 2 and 6 chains is delayed in the medium supplemented with FCS, and the 6 chain is absent in the medium supplemented with Ultroser G; when they are confluent or blocked by mitomycin C, greater changes are observed only in the medium supplemented with Ultroser G and lead to inhibition or delay of the expression of 2 and 6. In the presence of fibroblasts, only the expression of filaggrin (marker of terminal differentiation) is affected; it is delayed in the medium supplemented with FCS whatever the state of fibroblasts, and is inhibited in the medium supplemented with Ultroser G in the presence of proliferating and confluent fibroblasts.Abbreviations DMEM
Dulbecco's Modified Eagle's Medium
- EGF
epidermal growth factor
- K-SMM
keratinocyte-serum free medium
- PBS
phosphate-buffered saline Ca2+- and Mg2-free 相似文献
9.
In the course of characterization of glycolipid sulfotransferase from human renal cancer cells, the manner of inhibition of sulfotransferase activity with pyridoxal 5-phosphate was investigated. Incubation of a partially purified sulfotransferase preparation with pyridoxal 5-phosphate followed by reduction with NaBH4 resulted in an irreversible inactivation of the enzyme. When adenosine 3-phosphate 5-phosphosulfate was co-incubated with pyridoxal 5-phosphate, the enzyme was protected against this inactivation. Furthermore, pyridoxal 5-phosphate was found to behave as a competitive inhibitor with respect to adenosine 3-phosphate 5-phosphosulfate with aK
i value of 287 µm. These results suggest that pyridoxal 5-phosphate modified a lysine residue in the adenosine 3-phosphate 5-phosphosulfate-recognizing site of the sulfotransferase. 相似文献
10.
We have compared carbonyl diimidazole (CDI) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC) as activating agents for the oligomerization of negatively-charged - and -amino acids in homogeneous aqueous solution. -Amino acids can be oligomerized efficiently using CDI, but not by EDAC. -Amino acids can be oligomerized efficiently using EDAC, but not by CDI. Aspartic acid, an - and -dicarboxylic acid is oligomerized efficiently by both reagents. These results are explained in terms of the mechanisms of the reactions, and their relevance to prebiotic chemistry is discussed. 相似文献
11.
Genotype-independent leaf disc transformation of potato (Solanum tuberosum) using Agrobacterium tumefaciens 总被引:1,自引:0,他引:1
M. De Block 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1988,76(5):767-774
Summary Leaves of the in vitro grown potato cultivars Bintje, Berolina, Desiree, and Russet Burbank were wounded and co-cultivated with Agrobacterium strains having chimeric bar and nptII genes on a disarmed T-DNA. Each leaf from these cultivars formed numerous calli on kanamycin-containing medium, and almost all calli regenerated shoots. For Russet Burbank, it was necessary to include AgNO3 in the medium to obtain efficient shoot regeneration. The transformed plants have one to a few copies of the T-DNA, show NPT-II and PAT activities, and are resistant to high doses of the commercial preparation of phospinotricin (glufosinate). Almost no somaclonal variation was detected in trans-genic plants. 相似文献
12.
Rajendra Prasad Roy Parimala Nacharaju Ronald L. Nagel A. Seetharama Acharya 《Journal of Protein Chemistry》1995,14(2):81-88
Interspecies hybrids of HbA and Hb from mouse C57BL/10 [
2
M
2
H
and
2
H
2
M
(H=human, M=mouse)], representing 19 and 27 sequence differences per dimers (as compared with human dimer) have been generatedin vitro. The efficiency of the assembly of the interspecies hybrids by the alloplex intermediate pathway is about twofold higher than the low-pH-mediated subunit approach. The interspecies hybrids exhibit a cooperative O2 binding. The intrinsic O2 affinity of mouse Hb is slightly lower than HbA, while the 2,3-diphosphoglycerate (DPG) effect is comparable. Interestingly, the interspecies hybrid
2
M
2
H
has high O2 affinity (compared to either human or mouse Hb), while the interspecies hybrid
2
H
2
M
exhibits a very low O2 affinity. These results suggest that the mouse chain generates a tetramer with very low oxygen affinity. However, the complementarity of the mouse and chains generates a set of unique interactions that compensate for the low-oxygen-affinity propensity of the mouse chain. DPG binds the tetramer in the central cavity formed by the two subunits, hence the DPG effects on the interspecies hybrids should be as in the parent molecule. However, the results of the present study demonstrate that the DPG binding pocket is influenced by the nature of the chain present in the tetramer. The mouse chain reduces considerably the DPG right shift of the O2 affinity of the human-chain containing hybrid. Sequence analysis suggest that perturbations of the
1
1 (not the
1
2) are communicated to the DPG binding pocket in the presence of the alien subunit, and are the primary determinant of the ligand binding properties. The results have implications for the design of Hb-based blood substitutes and understanding of the inhibitory potential of mouse chains in transgenic mouse expressing human
S chains. 相似文献
13.
A. M. Popov 《Journal of Evolutionary Biochemistry and Physiology》2003,39(3):314-320
Using permeability to labeled glucose as a criterion of stability for liposomal membranes, a comparative study on stabilizing properties of different sterols and triterpenes in phospholipid bilayer has been carried out as well as on structural peculiarities of sterols responsible for membranolytic properties of cucumarioside G1 from the cucumaria Eupentacta fraudatris. Stabilizing action of the studied sterols and triterpenoides incorporated in the bilayer decreases in the following order: cholesterol sulfate > cholesterol > 5-sterols > -sitosterol > ergosterols > 7-sterols > epicholesterol > pregnane > androstane > coprosterol > 14-methylcholest-9(11)-en-3-ol > 4, 14-dimethylcholest-9(11)-en-3-ol > holothurinogenin A1 > glucoside of cholesterol > -xylosidase of 7-sterols > betulin > protopanaxatriol > phosphatidylcholine liposomes without sterol > protopanaxadiol > oleanolic acid. Sterol-dependent membranolytic cucumarioside G1 practically loses its ability to increase permeability of phospholipid membranes containing sterols obtained from this holothuria as well as coprosterol, epicholestrol, sulfated and glycosylated forms of sterols. The obtained results confirm the sterol hypothesis of the mechanism of membranotropic action of holothuria glycosides and of resistance to them of holothuria cell membranes. 相似文献
14.
K. Chalvet-Monfray P. Auger L. P. Belzunces C. Fléché P. Sabatier 《Acta biotheoretica》1996,44(3-4):335-348
The aim of this work is to propose methods to test mechanism of synergy of toxic agents in bees. A synergy between prochloraz, an imidazole fungicide, and deltamethrin, a pyrethroid insecticide, was demonstrated experimentally. The hypothesis is that prochloraz modifies the penetration or the metabolism of deltamethrin. This hypothesis is tested using a pharmacokinetic box model. A previous experimental work showed that bee instantaneous mortalities were higher, from the time t
1 to the time t
2 after spraying, in groups sprayed with deltamethrin at dose D
0 in the presence of prochloraz (+P) than in those sprayed with deltamethrin alone at a dose time as high (). We postulate that accrued mortality is proportional to the cumulated internal deltamethrin (ID
2). ID
2 of treatment (+P) had to be greater than ID
2 of treatment () during the period from t
1 to t
2 so that the hypothesis would be consistent with the experimental data. The limit, for which the hypothesis is conceivable, is the ID
2() = ID
2(+P
) curve. We study, in particular, the asymptotic behaviour of the limit curve when different parameters of the kinetic model tend to 0 or . These limits allow to verify quickly and easily whether a mechanism is conceivable or not As the limits are calculated with algebraic values, the test can be used for other synergies. 相似文献
15.
N. Ito K. Nishi M. Nakajima Y. Okamura T. Hirota 《Histochemistry and cell biology》1988,89(2):121-128
Summary Effects of -galactosidase (from green coffee beans) digestion on lectin staining were examined in formalin-fixed, paraffin-embedded human pancreatic tissues from individuals of blood-group B and AB. Digestion with the enzyme resulted in almost complete loss of Griffonia simplicifolia agglutinin I-B4(GSAI-B4) staining in the acinar cells with concomitant appearance of Ulex europaeus agglutinin-I(UEA-I) staining in the corresponding cells. In addition, reactivity with soybean agglutinin(SBA) was also imparted by the enzyme digestion in GSAI-B4 positive acinar cells. -Galactosidase digestion following -galactosidase digestion neither reduced the reactivity with SBA nor induced the reactivity with Griffonia simplicifolia agglutinin-II(GSA-II) in GSAI-B4 positive cells, while in UEA-I positive cells, both reduction of SBA reactivity and appearance of GSA-II reactivity occurred after simple -galactosidase digestion as well as sequential digestion with - and -galactosidase. However, when -l-fucosidase digestion procedure was inserted between - and -galactosidase digestion, UEA-I staining imparted by -galactosidase digestion was markedly decreased in intensity and GSA-II reactivity was appeared in GSAI-B4 positive acinar cells. Furthermore, after sequential digestion with -galactosidase and fucosidase, reactivity with peanut agglutinin(PNA) was revealed in GSAI-B4 positive acinar cells as well as UEA-I positive cells in secretors. In non-secretors, strong PNA staining was usually observed in the acinar cells throughout the glands without enzyme digestion. These results confirmed that the -galactosidase induced GSA-II reactivity and the fucosidase induced PNA reactivity are due to precursors of different kinds of blood-group determinants and suggest that at least two kinds of B antigen determinants, i.e. Gal(1-3)[Fuc(1-2)]Gal(1-3,4)GlcNac and Gal(1-3)-[Fuc(1-2)]Gal(1-3)GalNAc are produced in GSAI-B4 positive acinar cells. The synthesis of the latter type of B antigen is assumed to be controlled under the secretory gene in human pancreas.Abbreviation GalNAc
N-acetyl-d-galactosamine
- Gal
d-galactose
- GlcNAc
N-acetyl-d-glucosamine
- Fuc
l-fucose
- NeuNAc
N-acetylneuraminic acid (sialic acid) 相似文献
16.
Mammalian brain tubulin is an heterodimer; both and exist in 6–7 isotypic forms which differ in their amino acid sequences. By the use of isotype-specific monoclonal antibodies, we have previously shown that we can purify the
II,
III, anda
IV tubulin dimers from bovine brain. We have also observed that these isotypes differ in their distributionin vivo and their polymerization and drug-binding propertiesin vitro. We have now explored the question of whether the isotypically purified dimers differ in their overall conformation using as probes compounds of theN,N-polymethylenebis (iodoacetamide) series which are known to form discrete intrachain cross-links in-tubulin. These compounds have the structure ICH2CONH(CH2)
n
NHCOCH2I. One of these cross-links, designated
s, is between cys12 and either cys201 or cys211. The other, designated
*, is between cys239 and cys354. The
* cross-link forms in
II and
IV but not in
III; this is not surprising in view of the fact that
III has serine at position 239 instead of cysteine. However,
III is also unable to form the
s cross-link, although it appears to have all three cysteines which may be involved in the cross-link. This suggests that at least one of the sulfhydryls involved in the cross-link may be inaccessible in
III. Although both
II and
IV can form the
s cross-link, the dependence on cross-linker chain length is different.
II forms
s with derivatives in whichn=2, 4, 5, 6, and 7 but not with those in whichn=3 or 10. In contrast,
IV forms
s with derivatives in whichn=2, 3, 4, 5, 6, 7, and 10. These results imply that the
s sulfhydryls are slightly more accessible in
IV and are therefore less dependent on the conformation of the cross-linker to react with it. It appears, therefore, that the
II,
III, and
IV dimers each have unique conformations. This may help to explain the different assembly and drug-binding properties of these dimers. 相似文献
17.
Bennett T. Farmer II Luciano Müller Edward P. Nikonowicz Arthur Pardi 《Journal of biomolecular NMR》1994,4(1):129-133
Summary A set of three 3D (1H, 13C, 15N) triple-resonance correlation experiments has been designed to provide H1-H8 intraresidue sugar-to-base correlations in purines in an unambiguous and efficient manner. Together, the HsCsNb, HsCs(N)bCb, and HbNbCb experiments correlate the H1 sugar proton to the H8 proton of the attached base by means of the {H1, C1, N9, C8, H8} heteronuclear scalar coupling network. The assignment strategy presented here allows for unambiguous H1-H8 intraresidue correlations, provided that no two purines have both the same H1 and C1 chemical shifts and the same C8 and N9 chemical shifts. These experiments have yielded H1-H8 intraresidue sugar-to-base correlations for all five guanosines in the [13C, 15N] isotopically labeled RNA duplex r(GGCGCUUGCGUC)2. 相似文献
18.
R. Lohrmann 《Journal of molecular evolution》1977,10(2):137-154
Summary Adenosine 5-phosphoramidates form when solutions containing adenosine 5-polyphosphates pnA (n 3) or P1, P2-diadenosine 5-diphosphate and amines are allowed to dry out. Mg ions catalyze these reactions. We have studied systems containing ammonia, imidazole, glycine, ethylenediamine and histamine. The yields of adenosine 5-phosphoramidates range from 10–50 % based on the nucleotide. The prebiotic significance of the reactions is discussed.Abbreviations Im
imidazole
- hist
histamine
- gly
glycine
- en
ethylenediamine
- CDI
1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride
- EDTA
ethylenediaminetetraacetic acid
- A
adenosine
- Pn (n = 1, 2 )
linear polyphosphate containing n phosphate residues
- pnA
adenosine 5-polyphosphate containing n phosphate residues
- ADP
adenosine 5-diphosphate
- ATP
adenosine 5-triphosphate
- AppA
P1, P2-diadenosine 5-diphosphate
- gly-pA
adenylyl-(5N)-glycine
- ImpA
adenosine 5-phosphorimidazolide
- NH2-pA
adenosine 5-phosphoramidate
- en-pA
adenylyl-(5N)-ethylenediamine
- hist (NH) - pA
adenosine 5-phospho-[2-(4-imidazolyl)-ethylamide]
- hist(Im)-pA
adenosine 5-phospho-[4-(2-aminoethyl)-imidazolide]
- enP1,2
phosphoramidates of ethylenediamine derived from H3PO4 and H4P2O7 相似文献
19.
1. Opioid (, , ) and cannabinoid (CB1, CB2) receptors are coupled mainly toGi/Go GTP-binding proteins. The goal of the present study was to determine whether different subtypes of opioid and cannabinoid receptors, when coexpressed in the same cell, share a common reservoir, or utilize different pools, of G proteins.2. The stimulation of [35S]GTPS binding by selective opioid and cannabinoid agonists was tested in transiently transfected COS-7 cells, as well as in neuroblastoma cell lines. In COS-7 cells, cotransfection of - and -opioid receptors led to stimulation of [35S]GTPS binding by either -selective (DAMGO) or -selective (DPDPE) agonists. The combined effect of the two agonists was similar to the effect of either DAMGO or DPDPE alone, suggesting the activation of a common G-protein reservoir by the two receptor subtypes.3. The same phenomenon was observed when COS-7 cells were cotransfected with CB1 cannabinoid receptors and either - or -opioid receptors.4. On the other hand, in N18TG2 neuroblastoma cells, which endogenously coexpress CB1 and -opioid receptors, as well as in SK-N-SH neuroblastoma cells, which coexpress - and -opioid receptors, the combined effects of the various agonists (the selective cannabinoid DALN and the selective opioids DPDPE and DAMGO) were additive, implying the activation of different pools of G proteins by each receptorsubtype.5. These results suggest a fundamental difference between native and artificially transfected cells regarding the compartmentalization of receptors and GTP-binding proteins. 相似文献
20.
Luc Plateaux 《Insectes Sociaux》1960,7(3):221-226
Résumé Nous avons fait élever des larves d'Anergates atratulus par des ouvrières deMyrmica laevinodis à 22°C. Pour y parvenir, il n'est pas utile de faire hivernerensemble les larves d'Anergates et les ouvrières deMyrmica. La présence de larves autochtones n'empêche pas lesMyrmica d'élever des larves d'Anergates. Dans toutes les expériences lesMyrmica ont été soumises au fridavant de recevoir des larves d'Anergates. Aucune reine deMyrmica n'a été utilisée dans ces expériences.Sur les 64 larves d'Anergates que nous avons utilisées, 38 se sont transformées en imagos. C'est au début de l'adoption et au moment des métamorphoses que périrent la plupart des 26Anergates perdus. Les femelles vécurent en général 2 ou 3 jours et cherchèrent très tôt à quitter le nid natal. Les mâles vécurent 2 à 3 semaines.
Summary Larvae ofAnergates atratulus were experimentally reared by workers ofMyrmica laevinodis, at 22°C. An overwintering of both larvae ofAnergates and workers ofMyrmica is not necessary for the success of that experiment. The presence of larvae ofMyrmica does not keep theMyrmica from rearing larvae ofAnergates. The workers ofMyrmica have been cooled, in all the experiments, before receiving larvae ofAnergates. No queen ofMyrmica have been used in that experiments.38 of the 64 larvae ofAnergates used became imagos. Most of the 26 lostAnergates died at the beginning of the adoption and during the metamorphosis. The females lived generally 2 or 3 days and tried, very early, to leave their native nest. The males lived 2 or 3 weeks.
Anergates atratulus Myrmica laevinodis, 22 . bmecme Anergates Myrmica. Myrmica Anergates. Myrmica Anergates. Myrmica . 64 Anergates , 38 . 26 Anergates 2 3 . 2 3 .相似文献