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1.
The polypeptide structure and assembly of Ly-2/3 heterodimers 总被引:1,自引:0,他引:1
Mild reduction of mature, thymic Ly-2/3 heterodimers of M
r 67 000 resulted in dissociation into three individual polypeptide chains, , , and , of respective M
r values 38000, 35000, and 30000. The and chains were both immunoprecipitated by a monoclonal antibody directed to the Ly-2.1 epitope whereas the Ly-3.1 antibody bound only the chain. The possibility that the and chains of each heterodimer established their interchain links within a labile precursor protein in which a and segments were fused was considered but discounted by the finding that in mice heterozygous for both Ly-2 and Ly-3 loci, the Ly-2 product of one chromosome was not exclusively joined to Ly-3 structures coded by the same chromosome. By utilizing ionic detergents which selectively alter the charge of intrinsic membrane proteins, both Ly-2 and Ly-3 polypeptides were shown to have membrane insertion sites. It is suggested that as a consequence of their likely synthesis on membrane-bound polysomes, newly synthesized Ly-2 and Ly-3 structures accumulate within the same subcellular compartment — the membranes of the rough endoplasmic reticulum. Their elevated concentration within this space may facilitate a low affinity binding interaction between Ly-2 and Ly-3 which is later stabilized by interchain disulfide bond formation.Abbreviations used in this paper BSA
bovine serum albumin
- DOC
sodium deoxycholate
- DTT
dithiothreitol
- HA
hemagglutinin
- HTAB
hexadecyltrimethylammonium bromide
- RER
rough endoplasmic reticulum
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoresis
- TX100
Triton X-100 相似文献
2.
Jacques Raymond Brahim Mimouni Jean-Louis Azanza 《Plant Systematics and Evolution》1994,193(1-4):69-79
The seed storage globulins from sixHelianthus and four hybrids were studied using mono and bidimensional gel SDS electrophoresis (+ 2 mercaptoethanol). The polypeptide composition of each subunit was determined. Different pairs are specifically expressed according to the species studied. Three typical patterns were discriminated. All the studied species exhibit five subunits: two of them are expressed in all the species (11 and 22). The subunit corresponding to the 11 pair is present inH. petiolaris and in the three populations ofH. annuus studied. The 2b2 pair is common toH. annuus andH. argophyllus. H. petiolaris presents two specific 2a2 and 44 pairs andH. annuus a specific 33 pair. InH. argophyllus 11 33 or 44 are never observed but are replaced by 13 and 31 pairs. Some globulins, poorly represented, are of forms but present chains of higher molecular weights (in the range 54–56 kDa). Expressing variations in the banding patterns between these species by the use of a similarity index reveals complete identity between the three populations ofH. annuus. Identity between the twoH. petiolaris studied is also observed.H. annuus andH. argophyllus appear to be closer to each other thanH. petiolaris concerning the seed storage globulins. 相似文献
3.
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
4.
J. Romero-Andreas F. A. Bliss 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,71(3):478-480
Summary Crude protein extracts from single seeds of nondomesticated Mexican bean accessions were analysed by SDS polyacrylamide gel electrophoresis for variability in phaseolin protein. Six new phaseolin types; M1, M2, M3, M4, M5, M6, which contained polypeptides within the same range of molecular weights (51,000 to 45,000 daltons) as occur in the S, T and C phaseolin types of cultivated beans were identified. No T and C types were found among the non-domesticated Mexican accessions, and the S type occurred in less than 7% of the seeds screened. Genetic analyses of F2 progenies from crosses between Sanilac (S), and five of the M types showed that each M phaseolin phenotype was allelic to the S type and expressed codominantly. 相似文献
5.
The role of the subunits of phosphoinositide (PI) 3-kinase in NF-B activation in silica-stimulated RAW 264.7 cells was investigated. Results indicate that PI3-kinase activity was increased in response to silica. The p85 subunit of PI3-kinase interacted with tyrosine-phosphorylated IB- in silica-stimulated cells. PI3-kinase specific inhibitors, such as wortmannin and LY294003, substantially blocked both silica-induced PI3-kinase and NF-B activation. The inhibition of NF-B activation by PI3-kinase inhibitors was also observed in pervanadate-stimulated but not in LPS-stimulated cells. Furthermore, tyrosine phosphorylation of NF-B p65 was enhanced in cells stimulated with silica, pervanadate or LPS, and wortmannin substantially inhibited the phosphorylation event induced by the first two stimulants but not LPS. Antioxidants, such as superoxide dismutase (SOD), N-acetylcysteine (NAC) and pyrrolidine dithiocarbamate (PDTC), blocked silica-induced PI3-kinase activation, suggesting that reactive oxygen species may be important regulatory molecules in NF-B activation by mediating PI3-kinase activation. Our data suggest that p85 and p110 subunits of PI3-kinase play a role in NF-B activation through interaction with tyrosine-phosphorylated IB- and contributing to tyrosine phosphorylation of p65 NF-B. 相似文献
6.
Eva Halapi Yasuyoshi Yamamoto Claes Juhlin Mahmood Jeddi-Tehrani Johan Grunewald Roland Andersson Chistina Hising Giuseppe Masucci Håkan Mellstedt Rolf Kiessling 《Cancer immunology, immunotherapy : CII》1993,36(3):191-197
Tumour-infiltrating lymphocytes (TIL) are often observed in human tumours and their presence has been correlated with a better prognosis. It has been suggested that TIL are enriched for tumour-specific cytotoxic cells, and TIL activated and expanded in vitro by interleukin-2 (IL-2) are currently used in the therapy of human cancer. We have studied the T cell repertoire in IL-2-expanded TIL cells from patients with ovarian and renal carcinoma using T-cell-receptor-V--specific monoclonal antibodies and a polymerase-chain-reaction-based Southern blot technique for analysis of J- usage. In TIL lines derived from three of nine patients with ovarian carcinomas and from two of eight patients with renal carcinomas, selective usage of the V-6 or V-5 T-cell receptor gene products was found. The majority of the cells were CD4+, with up to 40% of the T cells utilizing the same V- gene. T-cell lines derived from peripheral blood lymphocytes from patients or healthy donors contained normal levels of V- subsets. Only moderate levels of V-6+ T cells were detected from freshly isolated TIL and the increase of this subpopulation appeared as a result of in vitro culture. The level of clonal restriction, as measured by the usage of J- gene segments within the V-5 or V-6 families, was analysed using a recently developed technique based on the polymerase chain reaction. Evidence for restricted J- usage was detected only in TIL expanded in vitro, while this was not the case in freshly isolated tumour-derived lymphocytes or T cell lines obtained from peripheral blood lymphocytes. The presence of a population with biased T cell receptor expression in cells derived from tumour tissue could be explained by their activation in vivo as a result of contact with tumour antigens and should be taken into consideration when discussing the therapeutic efficiency of IL-2-expanded TIL. 相似文献
7.
Purified ribulose-bisphosphate carboxylase (EC 4.1.1.39) was strongly and equally inhibited either by ADP or GDP and to a lesser extent by IDP. AMP or ATP exerted little effect on activity. Inhibition by the nucleotide diphosphates was competitive with respect to RuBP and non-competitive with respect to CO2 and Mg2+, respectively. Treatment of the enzyme with urea or guanidine-HCl resulted in rapid loss of activity that was not restored by dialysis even in the presence of Mg2+ and cysteine. Sodium dodecyl sulfate electrophoresis of 8.0 M urea treated enzyme revealed the presence of a fast-moving (small) sub-unit with molecular weight 14150 and a slower moving (large) sub-unit with molecular weight 68000. Examination of native enzyme by sodium dodecyl sulfate electrophoresis gave sub-units of 13700 and 55500 respectively. The amino acid content standardized to phenylalanine was essentially similar to that from other sources. Arrhenius plots showed a break at 29°C with an E
a of 12.34 kcal per mole for the steeper part of the curve and a H of 11.43 kcal per mole while for the less steep region, the E
a was 1.04 kcal per mole and the H 1.92 kcal per mole.Abbreviations ADP
adenosine-5-diphosphate
- AMP
adenosine-5-monophosphate
- ATP
adenosine-5-triphosphate
- CDP
cytidine-5-diphosphate
- CMP
cytidine-5-monophosphate
- CTP
cytidine-5-triphosphate
- FDP
fructose-1,6-diphosphate
- F6P
fructose-6-phosphate
- GDP
guanosine-5-diphosphate
- GMP
guanosine-5-monophosphate
- G6P
glucose-6-phosphate
- GTP
guanosine-5-triphosphate
- IDP
inosine-5-diphosphate
- IMP
inosine-5-monophosphate
- PEP
phosphoenolpyruvate
- 6PG
6-phosphogluconate
- R1P
ribose-1-phosphate
- R5P
ribose-5-phosphate
- RuBP
ribulose-1,5-bisphosphate
- SDS
sodium dodecyl sulfate
- TDP
thymidine-5-diphosphate
- TMP
thymidine-5-monophosphate
- TTP
thymidine-5-triphosphate
- UDP
uridine-5-diphosphate
- UMP
uridine-5-monophosphate
- UTP
uridine-5-triphosphate 相似文献
8.
S. -J. Yin W. F. Bosron T. -K. Li K. Ohnishi K. Okuda H. Ishii M. Tsuchiya 《Biochemical genetics》1984,22(1-2):169-180
Liver homogenate-supernatants from most Japanese exhibit an atypical pH optimum for ethanol oxidation at pH 8.8 instead of 10.5, the typical pH-activity optimum. It has been proposed that atypical livers contain alcohol dehydrogenase isozymes with 2 subunits while typical livers contain isozymes with 1 subunits, both produced by the ADH
2 gene. Because it is difficult to differentiate the atypical ADH2 2-2 phenotype from the ADH2 2-1 phenotype by starch gel electrophoresis, an agarose isoelectric focusing procedure was developed that clearly separated the atypical Japanese livers into two groups, A1 and A2. The isozymes in A1 and A2 livers were purified. Type A1 livers contained a single isozyme with an atypical pH-rate profile; it was designated 22. Three isozymes were isolated from A2 livers, two of which corresponded to 11 and 22. A third, absent from the typical and the atypical A1 livers, had an intermediate mobility; it was designated 21. Type A1 livers are, therefore, the homozygous ADH2 2-2 phenotype, and type A2 livers, the heterozygous ADH2 2-1 phenotype. The ADH2 2-2 phenotype was found in 53% of 194 Japanese livers, and the ADH2 2-1 phenotype, in 31%. Accordingly, the frequency of ADH
2
2
was 0.68.This study was supported by U.S. Public Health Service Grant AA 02342. 相似文献
9.
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
10.
Identification of a high-molecular-weight subunit of glutenin whose presence correlates with bread-making quality in wheats of related pedigree 总被引:12,自引:0,他引:12
P. I. Payne K. G. Corfield J. A. Blackman 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1979,55(3-4):153-159
Summary The subunit composition of glutenin was analysed by SDS-polyacrylamide-gel electrophoresis using two varieties of contrasting pedigrees. Maris Widgeon, a variety of good bread-making quality, was shown to contain 2 glutenin subunits not present in Maris Ranger, a much higher yielding variety that is unsuitable for making bread. A third subunit was only found in Maris Ranger glutenin. To determine if any of these subunits are directly related to bread-making quality, 60 randomly-derived F2 progeny from a Maris Widgeon x Maris Ranger cross were analysed for bread-making quality and for glutenin subunit composition. A strong correlation was demonstrated between the presence of one of the two subunits inherited from Maris Widgeon, and quality. This subunit (termed subunit 1 glutenin) had an approx. mol. wt. of 145,000. It was also found in Maris Freeman, a bread-making variety selected from the same cross previously made in 1962. In further crosses involving Maris Widgeon or its descendants, more bread-making varieties have been produced in the last decade at the Plant Breeding Institute, Cambridge and all but one have inherited glutenin subunit 1. The subunit has been traced back through Holdfast to White Fife, a Canadian hard spring wheat of excellent breadmaking quality. Some 67 varieties were screened for the presence of glutenin subunit 1 and it was found in 31% of them. Several unrelated varieties of good bread-making quality did not contain subunit 1 glutenin. 相似文献
11.
Decroocq V. Hagen L.S. Favé M.-G. Eyquard J.-P. Pierronnet A. 《Molecular breeding : new strategies in plant improvement》2004,13(2):135-142
In a previous study, cDNA microsatellite markers were described in apricot (Prunus armeniaca L.). Specific PCR primers were designed to amplify the microsatellite-containing regions from genomic DNA in different Prunus species. In the present work, cDNA microsatellite markers were developed in the hexaploid Prunus domestica L. species and polymorphism was ascertained in a segregating plum population. Co-dominant mendelian segregation of alleles was demonstrated and microsatellite polymorphism displayed up to 6 alleles per SSR locus per individual. Parentage lineage of three full-sib European plum cultivars (cv. Cacanska najbolja, Cacanska rana and Cacanska lepotica) was reconstructed by the analysis of the above nuclear SSR markers, completed by four chloroplastic microsatellite loci. The six most informative nuclear loci enabled discrimination between the three Cacak cultivars and unrelated individuals as well as the previously proposed parents, Wangenheim and Pozegaca. Data obtained support previous evidence that these cultivars originated from the Stanley cultivar. However, SSR analysis finally excluded Wangenheim as the other possible parent. Based on the results obtained with nuclear and chloroplast SSR loci, we propose the origin of those three Cacak cultivars in a cross between Stanley as the mother plant and Ruth gerstetter as the pollinator. Furthermore, we demonstrate the utility of these apricot SSR markers for genotype fingerprinting of the hexaploid plum cultivars. 相似文献
12.
Lin CC Sun CC Luo SF Tsai AC Chien CS Hsiao LD Lee CW Hsieh JT Yang CM 《Journal of biomedical science》2004,11(3):377-390
Interleukin-1 (IL-1) has been recognized as a potent stimulus for the synthesis of prostaglandin (PG), which has been implicated in inflammatory responses of the airways. However, the mechanisms underlying IL-1-induced cyclooxygenase (COX) expression and PGE2 synthesis via activation of p42/p44 and p38 mitogen-activated protein kinases (MAPKs) in human tracheal smooth muscle cells (HTSMCs) are not completely understood. We found that IL-1 increased COX-2 expression and PGE2 synthesis in time- and concentration-dependent manners. Both specific phosphatidylcholine-phospholipase C inhibitor (D609) and protein kinase C inhibitor (GF109203X) attenuated IL-1-induced responses in HTSMCs. IL-1-induced COX-2 expression and PGE2 synthesis were also inhibited by an inhibitor of MEK1/2 (PD98059) and inhibitors of p38 MAPK (SB203580 and SB202190), respectively, suggesting the involvement of p42/p44 and p38 MAPKs in these responses. This hypothesis was further supported by the transient activation of p42/p44 and p38 MAPKs induced by IL-1. Furthermore, IL-1-induced activation of nuclear factor-B (NF-B) was inversely correlated with the degradation of IB- in HTSMCs. IL-1-induced COX-2 expression and PGE2 synthesis were inhibited by the NF-B inhibitor pyrrolidinedithiocarbamate. These findings suggest that the expression of COX-2 is correlated with the release of PGE2 from IL-1-challenged HTSMCs, which is mediated, at least in part, through p42/p44 and p38 MAPKs and NF-B signaling pathways in HTSMCs. 相似文献
13.
Hubert Felle 《Planta》1981,152(6):505-512
In the aquatic liverwort Riccia fluitans, membrane depolarization (m), change in membrane conductance (gm), and current-voltage (I-V) characteristics in the presence of different amino acids as well as the uptake of 14C-labeled amino acids were measured. L-isomers of the tested amino acids generate larger electrical effects (m, gm) than D-isomers, and the I-V characteristics show that the positive electrical inward-current of 20 mA m-2 generated by 0.5 mM D-serine is only about 50% of the current generated by adding 0.5 mM L-serine. Whereas - and -amino acids rapidly depolarize the membrane to the same extend, with -aminobutyric acid (-AB) and dipeptides no significant electrical effects have been measured. The uptake kinetics of 14C-labeled amino acids display three components: (I) A saturable high-affinity component with Ks-values of 48 M D-alanine, 12 M -aminoisobutyric acid (AIB), 9 M L-alanine, 8 M L-proline, and 6 M L-serine, respectively; (2) an apparently linear low-affinity component, and (3) an also linear but unspecific component at concentrations >20 times the given Ks-value. Uptake of 14C-labeled AIB can be inhibited competitively by all tested neutral amino acids, the L-isomers being more effective than the D-isomers, as well as by ammonium or methylamine. Vice versa, AIB competitively inhibits uptake of L-serine and L-alanine. It is concluded that an uncharged stereospecific carrier for the investigated amino acids exists in the plasmalemma of Riccia fluitans. Accumulation ratios of about 50 suggest secondary active transport driven by a transmembrane electro-chemical gradient (mainly m) which is generated by the electrogenic proton pump. It is suggested that this carrier binds to the amino group forming either a charged binary complex with positively charged amines (Felle 1980), or an uncharged complex with -AB or dipeptides, whereas electrogenic transport of - and -amino acids is mediated by a ternary carrier complex, probably charged by a proton.Symbols and Abbreviations m
membrane potential (mV)
- Eco
equilibrium potential (mV) of the transport system
- gm
membrane (slope) conductance (Sm-2)
- gm
change in gm
- I-V curve
current-voltage curve
- AIB
-aminoisobutytric acid
- -AB
-aminobutyric acid 相似文献
14.
This paper describes a model of a neural visual system of a higher animal, in which the capability of pattern recognition develops adaptively. To produce the adaptability, we adopted self-organizing cells, and with them modeled feature-detecting cells which were discovered by Hubel and Wiesel and whose plasticity was found by Blakemore and Cooper. Combining the self-organizing cells and the learning principle of a Perceptron-type system, we constructed a model of the whole visual system. The model is also equipped with an eye movement control mechanism for gazing, which reduces the number of selforganizing cells required for pattern recognition, thus contributing to their quick self-organization. Computer simulation and an experiment using a hardware simulator showed that self-organizing cells quickly become sensitive to the features often seen and that the resulted system can classify patterns with a rather small number of feature-detecting cells. 相似文献
15.
Cell wall polysaccharides in developing barley coleoptiles were examined using acetic acid–nitric acid extraction, alditol acetate and methylation analyses and enzymatic digestion. The coleoptile cell wall from imbibed grain was rich in pectic polysaccharides (30 mol%), arabinoxylan (25 mol%), cellulose (25 mol%) and xyloglucan (6 mol%), but contained only low levels of (13,14)--D-glucan (1 mol%). During 5 days of coleoptile growth, pectic polysaccharides decreased steadily to about 9 mol%, while (13,14)--D-glucan increased to 10 mol%. Following the cessation of growth of the coleoptiles at about 5 days, (13,14)--D-glucan content rapidly decreased to 1 mol%. The cellulose content of the walls remained at about 35–40 mol% throughout coleoptile growth. Similarly, arabinoxylan content remained essentially constant at 25–30 mol% during growth, although the ratio of substituted to unsubstituted 4-linked xylosyl units decreased from about 4:1 to 1:1. Xyloglucan content ranged from 6 mol% to 10 mol% and the oligosaccharide profile determined using a xyloglucan-specific endoglucanase and MALDI-TOF mass spectrometry indicated that the oligosaccharides XXGG and XXGGG were the principal components, with one and two acetyl groups, respectively, Thus, dramatic changes in wall composition were detected during the growth of barley coleoptiles, both with respect to the relative abundance of individual wall constituents and to the fine structure of the arabinoxylans. 相似文献
16.
The main polysaccharide component of the thickened cell walls in the storage parenchyma of Lupinus angustifolius L. cotyledons is a linear (1 4)--linked d-galactan, which is mobilised after germination (L.A. Crawshaw and J.S.G Reid, 1984, Planta 160, 449–454). The isolation from the germinated cotyledons of a -d-galactosidase or exo-(1 4)--d-galactanase with a high specificity for the lupin galactan is described. The enzyme, purified using diethylaminoethyl-cellulose, carboxymethyl-cellulose and affinity chromatography on lactose-agarose, gave two bands (major 60 kDa, minor 45 kDa) on sodium dodecyl sulphate-gel electrophoresis, and two similar bands on isoelectric focusing (major, pI 7.0, minor pI 6.7, both apparently possessing enzyme activity). The minor component cross-reacted with an antiserum raised against, and affinity-purified on, the major band. Both components had a common N-terminal sequence. The minor component was probably a degradation product of the major one. The enzyme had limited -galactosidase action, catalysing the hydrolysis of p-nitrophenyl--d-galactopyranoside and (1 4)- and (1 6)--linked galactobioses. Lactose [-d-galactopyranosyl-(1 4)-d-glucose] was hydrolysed only very slowly and methyl--d-galactopyranoside not at all. Lupin galactan was hydrolysed rapidly and extensively to galactose, whereas other cell-wall polysaccharides (xyloglucan and arabinogalactan) with terminal non-reducing -d-galactopyranosyl residues were not substrates. A linear (1 4)--linked galactopentaose was hydrolysed efficiently to the tetraose plus galactose, but further sequential removals of galactose to give the tetraose and lower homologues occurred more slowly. Galactose, -galactonolactone and Cu+2 were inhibitory. No endo--d-galactanase activity was detected in lupin cotyledonary extracts, whereas exo-galactanase activity varied pari passu with galactan mobilisation. Exo-galactanase protein was detected, by Western immunoblotting of cotyledon extracts, just before the activity could be assayed and then increased and decreased in step with the enzyme activity. The exo-galactanase is clearly a key enzyme in galactan mobilisation and may be the sole activity involved in depolymerising the dominant (1 4)--galactan component of the cell wall.Abbreviations CM
carboxymethyl
- DEAE
diethylaminoethyl
- SDS-PAGE
sodium dodecyl sulphate-polyacrylamide gel electrophoresis
- TLC
thin-layer chromatography
We wish to thank CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) for the award of a studentship to M.S. Buckeridge, and the Government of São Paulo State, Brazil for granting him leave of absence. We are grateful to Dr. Amanda Heyller (Unilever Research Laboratory, Colworth House, Bedford, UK) for N-terminal sequence determinations, to Dr. Stuart Wilson (Stirling) for preparing gelatin SDS-gels and to Cristina Fanutti (Stirling) for purifying the xyloglucan oligosaccharide. 相似文献
17.
The present paper reports our attempts to determine whether the inclusion of 0.0014 mM Zn++ within a hydroponic culture medium affects the ability of 12-day-old Zea mays, cv. SS-522 to take-up [3H]-aflatoxin B1. Data from the corollary experiment, i.e., whether inclusion of aflatoxin affects the ability of Zea mays, cvs. Truckers White, X-Sweet and Merit to take-up 65ZnCl2 are presented also. This report is a preliminary to one regarding an in-progress analysis of whether pollutant levels of Zn++ affect aflatoxin uptake and distribution. In the absence of irrigating seedlings, which were grown in Perlite containing 65ZnCl2, with a solution containing mixed aflatoxins, the stem contained the greatest amount of label with root plus seed the next highest and the leaf the least for each of the cvs. In contrast, when the seedlings were irrigated with a solution containing mixed aflatoxins, the root plus seed contained either an amount nearly identical to (cv. Truckers White) or in excess of that within the stem (cvs. X-Sweet and Merit). Calculation of the percentages of aflatoxin-induced diminutions in leaf, stem and root label suggested that the aflatoxins interfered with the translocation of 65ZnCl2 from the root to the stem and leaf, at least for cvs X-Sweet and Merit. When 0.0014mM Zn++ as ZnSO4 was added to an incubation medium in which 12-day-old seedlings were suspended and plant growth assessed over 72 hours, a 15% increase (significant at 0.05 level) in seedling height over that of Zn++-deficient plants was observed. No differences in [3H]-aflatoxin B1 uptake were noted between those seedlings which were grown in either Zn++-containing or lacking media. Less than one % of the[3H]-aflatoxin B1 which was taken-up was recovered within chloroform extracts of the seedlings. The distributions of radioactivity from [3H]-aflatoxin B1 for leaf, stem, seed and root were 0, 57, 26 and 19% and 0, 26, 58 and 18% for Zn++-containing and -lacking media, respectively. 相似文献
18.
Characterization and gene organization of Taiwan banded krait (Bungarus multicinctus) gamma-bungarotoxin 总被引:1,自引:0,他引:1
-Bungarotoxin was isolated from Bungarus multicinctus (Taiwan banded krait) venom using a combination of chromatography on a SP-Sephadex C-25 column and a reverse-phase high-performance liquid chromatography column. Circular dichroism (CD) measurement revealed that its secondary structure was dominant with -sheet structure as is that of snake venom -neurotoxins and cardiotoxins. -Bungarotoxin exhibits activity on inhibiting the binding of [3H]quinuclidinyl benzilate to the M2 muscarinic acetylcholine receptor subtype, and competes weakly with radioiodinated -bungarotoxin for binding to the Torpedo nicotinic acetylcholine receptor. Moreover, the toxin inhibits collagen-induced platelet aggregation, with an IC50 of approximately 200 nM. The genomic DNA encoding the -bungarotoxin precursor is amplified by polymerase chain reaction (PCR). The gene is organized with three exons separated by two introns, and shares virtually identical overall organization with those reported for -neurotoxin and cardiotoxin genes, including similar intron insertions. The intron sequences of these genes share sequence identity up to 85%, but the exon sequences are highly variable. These observations suggest that -bungarotoxin, -neurotoxins, and cardiotoxins originate from a common ancestor, and the evolution of these genes shows a tendency to diversify the functions of snake venom proteins. 相似文献
19.
Hypoxia has been shown to activate the pleiotropic cytokine TNF in the lung. TNF in turn, is known to induce pulmonary vasoconstriction. Additional effects of this cytokine in hypoxia mediated cardiopulmonary remodeling are poorly understood. To further evaluate the role of TNF in chronic hypoxia we exposed TNF null (TNF–/–) and wild-type mice to three weeks of hypobaric hypoxia (10% O2). Equivalent erythocytosis (Hematocrit increased by 40%) developed in both genetic backgrounds. In contrast, right ventricular systolic pressure increased in response to three weeks of hypoxia in the wild-type mice ( 75%), yet was unaltered in the TNF–/– mice. Concomitantly right ventricular hypertrophy was attenuated in the TNF–/– mice (35 ± 5% increase) when compared to wild-type mice (124 ± 6% increase p < 0.001, n 20). Interestingly in both strains the lung wet weights increased to a similar degree in response to hypoxia. In conclusion, our data demonstrate that TNF is an integral autocoid in chronic hypoxia mediated right ventricular hypertrophy. Moreover, additional components of cardiopulmonary remodeling may be regulated by TNF signaling as suggested by the negligible right ventricular systolic pressure response to hypoxia in the absence of TNF. 相似文献
20.
L. Goldman 《European biophysics journal : EBJ》1989,16(6):321-325
The time constant of the process producing the delay in Na inactivation development as determined by the two pulse method (delay) was extracted and compared to that of the slowest Na activation process 3 for the I
Na during the conditioning pulse of that same determination. delay and two pulse inactivation c values were computer generated using a nonlinear least squares algorithm. h and single pulse inactivation h values were independently generated for each determination also with the aid of the computer using the same non-linear least squares algorithm. In one determination at 2 mV, c was 4.68 and delay 0.494 ms while h was 4.70 and 3 0.491 ms for a c/h of 0.996 and a delay/3 of 1.006. Mean delay/3 from five determinations in four axons, both Cs and K perfused, and spanning a potential range of-27 to 2mV was 1.068. The precursor process to inactivation is channel opening. Some fraction of channels presumably inactivate via another route where prior channel opening is not required. 相似文献