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PubMed2004

Experimental and natural weed host-virus relations.

Weeds, as alternative hosts of plant viruses and nutrient plants of virus vectors play important role in virus ecology and epidemiology. The aim of our study was to discover new weed-virus relations. Therefore some weed species were mechanically inoculated with 28 viruses (strains or isolates) maintained in our glasshouse. Different weed species with and without visible symptoms were collected from agro-, water ecosystems and wastelands of Hungary between 1997 and 2003. Virus infections were evaluated by biotests, DAS ELISA serological methods, electronmicroscopy and immunosorbent electronmicroscopy (ISEM). Under glasshouse conditions Ambrosia artemisifolia was considered as a virophob species, showing resistance to all viruses listed above. A series of new artificial (Chenopodium album--SoMV (LH+SH)*, AMV (LH+SH); C. berlandieri--PVY(NTN) (LH), AMV (LH+SH), CMV (LH), SoMV (LH+SH), ObPV (LH+SH), ZYMV-10 (LH): C. ugandae--ObPV (LH), SoMV (L); C. glaucum--ObPV (LH), SoMV (L); Echinocystis lobata--PVX (L), ZYMV (LH+SH); Solanum nigrum--MYFV (LH+SH), PVY(N) (L), PVY(NTN) (LH+SH), SoMV (LH), TMV (SH), CMV (SH); S. dulcamara--CMV-U/246 (SH), PVY(NTN) (LH), SoMV-H (L), TMV-O (L); S. luteum--PVY(N) (SH), PVY(NTN) (LH+L), TMV(SH).) and natural (Asclepias syriaca--TMV, AMV, TSWV; Alisma plantago-aquatica--PVY, SoMV; Ambrosia artemisiifolia--CMV; Chenopodium album--CMV, PVS, PLRV; C. hybridum--CMV; Cirsium canum--CMV, PVM; Carex vulpina--CMV; Comium maculatum--PVY; Datura stramonium--PVA, PVX, PVS, PVM, CMV, TMV; Lysimachia vulgaris--ArMV, BNYVV, CMV, TMV; Lythrum salicaria--ArMV; Malva neglecta--CMV; Mercurialis annua--SoMV; Solanum nigrum--CMV, PVY, PVY(N); Solidago gigantea--CMV, RpRSV, BNYVV; Stenactis annua--PVM, PVA) weed--virus relations were detected. The epidemiological role of perennial hosts (A. syriaca, A. planlago aquatica, C. canurm, L. vulgaris, L. salicaria, S. gigantea) is especially high, because they can serve as infection sources as well as overwintering hosts of different plant viruses.

Antiviral Agents↗
PubMed1992

Structures of the asparagine-linked oligosaccharide chains of human von Willebrand factor. Occurrence of blood group A, B, and H(O) structures.

The asparagine-linked oligosaccharide chains of human von Willebrand factor (vWF) purified from pooled plasma were quantitatively liberated from the polypeptide moiety by hydrazinolysis. After N-acetylation, these were fractionated by paper electrophoresis and sequential chromatography on lectin-affinity columns of concanavalin A, Phaseolus vulgaris erythrophytohemagglutinin, Datura stramonium agglutinin, Ricinus communis agglutinin 120, and Ulex europaeus agglutinin I and on a Bio-Gel P-4 column. Their structures were investigated by sequential exoglycosidase digestion in conjunction with methylation analysis. The glycoprotein was shown to be unique in its great diversity of oligosaccharide structures. Another noteworthy finding which had not been reported previously was the occurrence of asparagine-linked oligosaccharide chains with blood group A, B, and H(O) structures. In the present study, this glycoprotein was shown to contain mono- (0.4% of the total oligosaccharides), bi-(78.2%), tri- (12.3%), and tetraantennary (2.3%) complex type oligosaccharides in addition to a series of high mannose type oligosaccharides, Man6-9GlcNAc2 (0.8%). Biantennary complex type oligosaccharide chains were those with (8.2%) and without (70.0%) a bisecting GlcNAc residue and approximately 13.2%, 2.2%, and 0.4% of these contained blood group H(O), A, and B structures, respectively. The tri- and tetraantennary complex type chains were those with and without N-acetyllactosamine repeats, and about 13.0% of the triantennary chains without the N-acetyllactosamine repeat contained the blood group H(O) structure. Occurrence of these asparagine-linked oligosaccharides with blood group A and B structures suggest that the repeated use of factor VIII/vWF pooled concentrate for the treatment of hemophiliacs could result in the production of antibodies against vWF with a different blood group from that of the patient, and this development may be pathogenic.

ABO Blood-Group System↗
PubMed2005

[Allelopathy of different plants on wheat, cucumber and radish seedlings].

By means of bioassay in laboratory and field, this paper studied the allelopathy of 18 kinds of plants in Gansu Province on the seedlings of wheat, cucumber and radish. The results showed that the aqueous extract of the stems and leaves of Artemisia annua, Solanum nigrum and Datura stramonium had the strongest allelopathy on test receptor plants, and their synthetic inhibitory effect (SE) was 47.66%, 32.89% and 26.63%, respectively. The SE of Xanthium sibiricum, Portulaca oleraca, Cephalanoplos segetum, and Chenopodium album was 21.71%, 20.93%, 20.83% and 20.2%, respectively, while Vicia amoena (SE 3.5%), Setaria viridis (SE 2.2%), and Cymamchum chinense (SE 1.97%) had a weaker allelopathy. Chenopodium ambrosioides (SE - 1.03%), Polygonum caespitosum (SE - 1.63%) and Avena fatua (SE 5.33%) had no evident allelopathy, but Artemisia annua affected the seedling height and fresh weight of radish, cucumber, wheat and maize, with the SE being 54.07%, 38.46%, 33.35% and 20.88%, respectively. Artemisia annua had a 44.70% of SE on wheat growth, and thus, had a certain value to develop and use.

Cucumis sativus↗
PubMed2005

History of aerosol therapy: liquid nebulization to MDIs to DPIs.

Inhaled therapies have been used since ancient times and may have had their origins with the smoking of datura preparations in India 4,000 years ago. In the late 18th and in the 19th century, earthenware inhalers were popular for the inhalation of air drawn through infusions of plants and other ingredients. Atomizers and nebulizers were developed in the mid-1800s in France and were thought to be an outgrowth of the perfume industry as well as a response to the fashion of inhaling thermal waters at spas. Around the turn of the 20th century, combustible powders and cigarettes containing stramonium were popular for asthma and other lung complaints. Following the discovery of the utility of epinephrine for treating asthma, hand-bulb nebulizers were developed, as well as early compressor nebulizers. The marketing of the first pressurized metered-dose inhaler for epinephrine and isoproterenol, by Riker Laboratories in 1956, was a milestone in the development of inhaled drugs. There have been remarkable advances in the technology of devices and formulations for inhaled drugs in the past 50 years. These have been influenced greatly by scientific developments in several areas: theoretical modeling and indirect measures of lung deposition, particle sizing techniques and in vitro deposition studies, scintigraphic deposition studies, pharmacokinetics and pharmacodynamics, and the 1987 Montreal Protocol, which banned chlorofluorocarbon propellants. We are now in an era of rapid technologic progress in inhaled drug delivery and applications of aerosol science, with the use of the aerosolized route for drugs for systemic therapy and for gene replacement therapy, use of aerosolized antimicrobials and immunosuppressants, and interest in specific targeting of inhaled drugs.

Administration, Inhalation↗
PubMed2005

Drug abuse in Slovak Republic.

The drug abusing structure has dramatically changed since 1989. While in 1989 the sniffing of the fluid drugs represented 98% of the global drug abuse, the most abused drugs were: heroin, marijuana, cocaine, amphetamine and its derivatives. During last 10 years situation with drug abuse has changed. Currently the most abused drugs: cannabinoides, amphetamines. The plant drugs (Datura stramonium, hallucinogenic mushrooms Psilocybe semilanceata, nutmeg--the seed of Myristica fragrans) combined with the alcohol are popular among the young abusers. According to an analysis of the phone consultations in our Toxicological Information Centre (TIC) we found out, that the number of intoxications with the plant drugs has increased five times during the last year (comparing with the year 2000), because of their easy availability, low price and quick spreading of information.

Adolescent↗
PubMed1991

Lectins from Triticum vulgaris and Limax flavus are universal antagonists of botulinum neurotoxin and tetanus toxin.

Lectins from Anguilla anguilla, Artocarpus integrifolia, Canavalia ensiformis, Datora stramonium, Glycine max, Limax flavus, Ricinus communis and Triticum vulgaris were tested for their abilities to antagonize the binding of botulinum neurotoxin and tetanus toxin to rat brain membranes and to antagonize the ability of these toxins to block neuromuscular transmission in mouse phrenic nerve-hemidiaphragm preparations. Lectins from Limax flavus and Triticum vulgaris, both of which have affinity for sialic acid, were antagonists of the various serotypes of botulinum neurotoxin and tetanus toxin. When tested against the high affinity binding site for botulinum neurotoxin type B, the lectin from Limax flavus had a Ki of 3.1 x 10(-7) M and the lectin from Triticum vulgaris had a Ki of 3.75 x 10(-7) M. When tested against the high affinity binding site for tetanus toxin, the lectins from Limax flavus and Triticum vulgaris had Ki values of 1.5 x 10(-7) and 1 x 10(-6) M, respectively. In all cases the lectins behaved as competitive antagonists. In reverse experiments, neither botulinum toxin nor tetanus toxin was a very effective antagonist of lectin binding to brain membranes. Studies on isolated neuromuscular preparations showed that the lectin from Triticum vulgaris did not affect transmission at concentrations of 10(-6) to 10(-3) M, but at a concentration of 3 x 10(-5) M the lectin produced highly statistically significant antagonism of the neuromuscular blocking properties of botulinum neurotoxin types A, B, C, D, E and F as well as tetanus toxin. The lectin did not antagonize beta-bungarotoxin.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2005

Purification of potato virus X and preparation of its antiserum.

Potato virus X (PVX) isolated from the potato leaf and tuber samples which were collected from various fields in Damavand and Ardabil. The initial isolations of the virus were made from potato by mechanical inoculation on Gomphrena globosa L. and Chenopodium spp. that produce local lesion, and then it causes mosaic on Nicotiana spp. and Datura stramonium L. An isolate of the virus inoculated to Nicotiana glutinosa L. and it was maintained throughout the work. Sap from infected N. glutinosa was ineffective after dilution to 10-6, 10 minutes at 70 degrees and 10 weeks at room temperature. The virus was readily purified from infected leaves and the best protocol was Moreira & Jones 1980 than the other 2 methods of Fribourg 1975 and Shepard & Shalla 1972. Antisera were prepared against native, degraded proteins and micro precipitin test showed that both antisera had a 1/512 titer. Precipitin lines with D - Protein antiserum was better of the native protein antiserum in agar double diffusion test than treated with SDS. The isolate of the virus was not transmitted by none of 2 species of Cuscuta but transmitted from infected leaves to healthy plants with sap inoculation without using Carburandum. This isolate showed positive reaction with gamaglubulin in kate received from CIP centre.

Enzyme-Linked Immunosorbent Assay↗
PubMed2005

Determination of the races of potato virus X and its host rang in Karaj, Damavand and Ardabil, Iran.

Potato virus x (PVX) is found commonly in potato-growing areas, worldwide. It is an economically important virus which causes losses in tuber yield of approximate 5 to 15 percent. In a 2 year survey, potato leaf and tuber samples were collected from various fields in Damavand and Karaj. The initial isolations from potato were made by mechanical inoculation first to Gomphrena globosa L. and later to Dartura stramonium L. It was not transmitted by 2 species of Cuscuta but transmitted mechamically. The isolates were inoculated to Nicotiana glutinosa L. in which they were maintained throughout the work. Sap from infected N. glutinosa was ineffective after dilution to 10-6, after 10 minutes at 70 degrees C and after 10 weeks at room temperature. The virus had filamentous and slightly flexuous particles with a normal length of about 490-500 nm and 12 nm width. According to the symptoms, TIP results and serological comparisons, the compared isolates showed no difference and they belong to XN group. In order to estimate disease incidence, 773 tubers from Damavand area were tested and compared with that in Ardabil area. Disease incidence in Damavand ranged from 1.1-20.9 percent and was lower than disease incidence in Ardabil. In 8 genera of collected weeds from fields of potato and tomato samples by using test plants and serological methods, they didn't show existence of the potato virus x.

Iran↗
PubMed2005

Weed control in sunflower (helianthus annuus L.) with post-emergent herbicides.

Sunflower is the most important oil crop in Hungary, is the base of the production of cooking oil and moreover takes an important part in production of margarine too. Extracted sunflower groats as a secondary product origining from the mentioned procedure can be used in forage successfully. The amount of harvested sunflower reaches the 20-25% of the EU's yield. The sowing area approaches 500 thousand hectares. The essential condition of successful crop production is the perfect weed control. Sowing areas are infected with monocotyledon and dicotyledonous weeds too. Annual dicotyledonous weeds are the most troublesome. The worst species is the Ambrosia artemisiifolia L. Many other weed species as Abutilon theophrasti MEDIC., Datura stramonium L. and Xanthium strumarium L. can cause serious damages. In our model experiments we examined the herbicide sensibility of two commercial sunflower cultivars as "Iregi szürke csikos", "Marica II" and a sulfonylurea-urea tolerant new hybrid "PR63E82". The experiment was set up under greenhouse conditions with the use of four important weed and different post-emergent herbicide as Modown 4F (bifenox), Pledge 50 WP (flumioxazin) and Granstar 75 DF (tribenuron-methyl). We applied normal and double doses too. Sunflower was cultivated to 4-6 leaf stage. Post-emergent herbicides were sprayed out when weeds were in 2, 2-4 and 4-6 leaf stage. Weed killer and phytotoxic effects of post-emergent herbicides were examined. We declared that development of weeds had significally effect on the effectiveness of different herbicides.

Adaptation, Physiological↗
PubMed1991

Carbohydrate-binding specificity of Tetracarpidium conophorum lectin.

The carbohydrate-binding specificity of a novel plant lectin isolated from the seeds of Tetracarpidium conophorum (Nigerian walnut) has been studied by quantitative hapten inhibition assays and by determining the behavior of a number of oligosaccharides and glycopeptides on lectin-Sepharose affinity columns. The Tetracarpidium lectin shows preference for simple, unbranched oligosaccharides containing a terminal Gal beta 1----4GlNAc sequence over a Gal beta 1----3GlcNAc sequence and substitution by sialic acid or fucose of the terminal galactose residue, the subterminal N-acetylglucosamine or more distally located sugar residues of oligosaccharides reduce binding activity. Branched complex-type glycans containing either Gal beta 1----4GlcNAc or Gal beta 1----3GlcNAc termini bind with higher affinity than simpler oligosaccharides. The lectin shows highest affinity for a tri-antennary glycan carrying Gal beta 1----4GlcNAc substituents on C-2 and C-4 of Man alpha 1----3 and C-2 of Man alpha 1----6 core residues. Bi- and tri-glycans lacking this branching pattern bind more weakly. Tetra-antennary glycans and mono- and di-branched hybrid-type glycans also bind weakly to the immobilized lectin. Therefore, Tetracarpidium lectin complements the binding specificities of well-known lectins such as Datura stramonium agglutinin, Phaseolus vulgaris agglutinin, and lentil lectin and will be a useful additional tool for the identification and separation of complex-type glycans.

Asialoglycoproteins↗
PubMed1991

[Mydriasis caused by plant contact].

Uni- or bilateral dilatation of pupils that are not reactive to light and lack miosis in response to 1% pilocarpine may be caused by contact with plants containing alkaloids such as scopolamine and atropine. Other causes of a non-light-reactive dilated pupil, such as Adie's tonic pupil, third nerve palsy and lesion of the mesencephalic pretectal region, must be excluded before testing the iris sphincter reaction to 1% pilocarpine. Among the naturally growing flowers in Germany, deadly nightshade (Atropa belladonna), jimson weed (thornapple, Datura stramonium) and black henbane (Hyoscyamus niger) contain enough alkaloids to cause mydriasis by direct contact. However, in most cases an accidental mydriasis by plants in Germany is caused by Datura arborea taxa, e.g. Datura suaveolens, D. candida, D. aurea and D. sanguinea. They contain up to 0.6% dry weight scopolamine. These plants can grow very large and are often planted in tubs. They have to be cut back each year before the winter. This is typically how the eye is contaminated by parts of the plants, which can cause dilatation of the pupil mimicing a neuroophthalmological disorder.

Diagnosis, Differential↗
PubMed1990

The polylactosaminoglycans of human lysosomal membrane glycoproteins lamp-1 and lamp-2. Localization on the peptide backbones.

Lysosome membrane glycoproteins, lamp-1 and lamp-2, have been shown to contain 18 and 16 N-glycans, some of which are modified by poly-N-acetyl-lactosamine. We have localized the polylactosaminoglycans to specific sites on lamp-1 and lamp-2 purified from human chronic myelogenous leukemia cells. Polylactosaminoglycan-containing glycopeptides, obtained by trypsin, pepsin, and V8 protease digestion of the glycoproteins, were isolated by Datura stramonium agglutinin affinity chromatography, gel filtration, and reverse phase high performance liquid chromatography. The poly-N-acetyllactosaminyl structures of isolated glycopeptides were confirmed by the susceptibility of their released oligosaccharides to endo-beta-galactosidase. Amino acid analysis and sequencing demonstrated that polylactosaminoglycans were located at Asn-34, Asn-93 and/or Asn-102, and Asn-195 and/or Asn-200 in lamp-1, and at Asn-4 and/or Asn-10, and Asn-279 in lamp-2. These results indicated that only certain glycosylation sites can be selectively modified by poly-N-acetyllactosamine, and those sites may confer the requirement by beta 1----3-N-acetylglucosaminyl transferase.

Amino Acid Sequence↗
PubMed1990

[Clinical effects of an extreme Ca overdose in feed for growing piglets].

In a piggery (approx. 60 sows) nearly all the weaned piglets (about 150) showed signs of poor development, reduced feed intake, increased water consumption, markedly dry and hard faeces, urine drinking, navel suckling and drowsiness over 6-8 weeks. In blood samples of 4 affected piglets a significant hypercalcaemia (values greater than 20 mg/dl)--combined with an at times marked hypophosphataemia--was measured. In the diet offered, a Ca content of 56.2 g/kg was estimated. An intoxication due to ochratoxin or due to excessive levels of quinoxaline or vitamin D was excluded by toxicological investigations of the diet. Contamination by Datura stramonium was also ruled out.

Animal Feed↗
PubMed1990

Glycoprotein storage in Gaucher disease: lectin histochemistry and biochemical studies.

Lectin histochemical studies were performed on formalin-fixed, frozen, and paraffin-embedded tissue sections from 19 patients with glucosylceramide lipidosis (i.e., Gaucher disease). Eleven different lectins were used to identify the specific carbohydrate residues in the undegraded stored compounds in the cytoplasm of Gaucher cells. In all cases studied, Gaucher cells stained with Concanavalia ensiformis agglutinin, Datura stramonium agglutinin, Lens culinaris, Ricinus communis agglutinin-I, and wheat germ agglutinin. These results demonstrated common carbohydrate residues in the undegraded material stored within Gaucher cells and indicated the presence of fucosylated N-linked complex oligosaccharides, and glycans containing N-acetyllactosamine repeating sequences, as well as nonreducing terminal beta-galactosyl and sialyl residues. In order to confirm these findings using biochemical methods, livers and spleens from Gaucher patients and controls, and from a patient with Niemann-Pick disease type C (included for comparison) were digested with Pronase and the resulting glycopeptides separated by gel filtration into fractions with high and low molecular weight. In the high-molecular-weight fractions from livers of Gaucher patients, the levels of sugars corresponding to N-linked glycans, as measured by gas-liquid chromatography, were elevated over those in controls. In the high-molecular-weight fractions from spleens, the levels of the same sugars were elevated in both Gaucher and Niemann-Pick type C patients. Digestion of the glycopeptides with endo-beta-galactosidase, which specifically cleaves polylactosaminoglycans, showed the presence of material containing N-acetyllactosamine repeating units in Gaucher liver glycopeptide fractions, but not in control and Niemann-Pick type C derived glycopeptide fractions. Our histochemical and biochemical studies demonstrated that in addition to glucosylceramide, affected tissues of patients with Gaucher disease accumulate glycoproteins. This accumulation could not have been predicted on the basis of the primary enzymatic defect.

Carbohydrates↗
PubMed1989

Decrease in polylactosaminoglycans associated with lysosomal membrane glycoproteins during differentiation of CaCo-2 human colonic adenocarcinoma cells.

The proportion of labeled polylactosaminoglycans found in glycoproteins decreases during spontaneous differentiation of CaCo-2 human colonic adenocarcinoma cells to enterocytes in culture (A. Youakim and A. Herscovics, Biochem. J., 247: 299-306, 1987). To identify polylactosaminoglycan-containing glycoproteins, CaCo-2 cells were incubated with [3H]glucosamine or [3H]fucose, for 24 h, and membrane glycoproteins solubilized with 0.5% Nonidet P-40 were fractionated by affinity chromatography on Datura stramonium (DSA)-agarose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography showed that a restricted set of glycoproteins with a molecular weight of about 100,000 bound to DSA-agarose. These labeled glycoproteins were shown to contain polylactosaminoglycans by DSA-agarose chromatography and endo-beta-galactosidase digestion of Pronase-derived glycopeptides. Immunoprecipitation of the [3H]glucosamine-labeled Nonidet P-40 extract with polyclonal antibodies to the lysosomal membrane proteins h-lamp-1 and h-lamp-2 followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography also revealed a band with a molecular weight of about 100,000. The immunoprecipitates were digested with Pronase, and the resulting glycopeptides were first fractionated on Bio-Gel P-6 into excluded (Fraction I) and included (Fraction II) glycopeptides, and then by DSA-agarose affinity chromatography. A much greater proportion of labeled glycopeptides in undifferentiated cells (3 to 5 days in culture) than in differentiated cells (19 to 27 days in culture) was recovered in Fraction I; these glycopeptides were bound to DSA-agarose and were sensitive to endo-beta-galactosidase. This decrease in polylactosaminoglycans was associated primarily with h-lamp-1. These results indicate that h-lamp-1 of CaCo-2 cells contains polylactosaminoglycans and that it undergoes a change in glycosylation with differentiation.

Adenocarcinoma↗
PubMed1988

Estrogen preferentially stimulates lactosaminoglycan-containing oligosaccharide synthesis in mouse uteri.

The effects of steroid hormones on the synthesis of lactosaminoglycan (LAG)-containing oligosaccharides by mouse uteri are reported. The uterine LAG-containing oligosaccharides were degraded partially by Pseudomonas endo-beta-galactosidase, releasing an oligosaccharide of the apparent structure: Gal beta----N-acetylglucosaminyl(----N-acetylgalactosaminyl)beta 1,3----galactose. A larger fraction of the LAG-containing oligosaccharides bound to pokeweed mitogen than to Datura stramonium lectin, suggesting the presence of highly branched structures. LAG-containing oligosaccharides were resistant to sequential digestion with Pronase, nitrous acid, hyaluronidase, and chondroitinase ABC. These polysaccharides exhibited a Gal:GlcNAc:GalNAc ratio of approximately 1.0:1.0:0.3 and were not fucosylated. The ion-exchange behavior of the LAG-containing oligosaccharides before and after mild acid hydrolysis indicated the presence of sialic acid residues. The LAG-containing glycopeptides were highly resistant to beta-elimination but were released quantitatively by hydrazinolysis, demonstrating an N-linkage to protein. Binding to pokeweed mitogen was markedly enhanced following release of these oligosaccharides from peptides by hydrazinolysis, suggesting that peptide-bound oligosaccharides were partially inaccessible to the lectin. Molecular exclusion chromatography of the oligosaccharides released by hydrazinolysis revealed a broad distribution ranging from Mr 4,000 to 15,000 with a median Mr of approximately 8,000. We extended the above observations by determining how the steroid hormones 17-beta-estradiol (E2) and progesterone affected synthesis of the LAG-containing oligosaccharides in ovariectomized mice. Generally, E2 and a number of E2 agonists stimulated glycoconjugate synthesis; however, chronic E2 treatment or combined treatment with E2 plus progesterone caused the synthesis of most glycosaminoglycans to return to basal levels. In contrast, E2 either alone or in combination with progesterone stimulated synthesis of LAG-containing oligosaccharides in preference not only to glycosaminoglycans but also to other classes of N-linked oligosaccharides. This effect was apparent during both priming and nidatory E2 treatments. Collectively, these data provide the first demonstration of LAG-containing oligosaccharides in uteri and for the hormonally regulated synthesis of lactosaminoglycans. In addition, this is the first demonstration of the ability of steroid hormones to induce the synthesis of certain types of N-linked oligosaccharides in preference to others in the same tissue.

Amino Sugars↗
PubMed1988

Purification and characterization of the D2-dopamine receptor from bovine anterior pituitary.

The D2-dopamine receptor from bovine anterior pituitary has been purified approximately 33,000-fold to apparent homogeneity by sequential use of affinity chromatography on immobilized carboxymethyleneoximinospiperone-Sepharose, Datura stramonium lectin-agarose, and hydroxylapatite chromatography. The purification yields a single polypeptide band of Mr approximately 120,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed by labeling with radioiodinated Bolton-Hunter reagent, Coomassie Blue, or silver staining. The purified D2 receptor preparations display a specific activity of approximately 5.3 nmol of [3H]spiperone bound per mg of protein. In detergent solutions, the purified receptor has a KD for [3H]spiperone of 5-8 nM; however, after reinsertion of the purified protein into phospholipid vesicles, a KD of approximately 160 pM is obtained, similar to that found for the receptor in crude membrane preparations. Several lines of evidence document that this polypeptide contains the ligand binding site as well as the functional activity of the D2 receptor. The Mr approximately 120,000 peptide can be covalently labeled by the affinity probe, 125I-bromoacetyl-N-(p-aminophenethyl)spiperone, with the pharmacological specificity expected of a D2-dopamine receptor. Agonist and antagonist ligands compete for [3H]spiperone binding to purified receptors in phospholipid vesicles with a rank order of potency and selectivity typical of a D2-dopamine receptor. Moreover, when reinserted into phospholipid vesicles with purified brain Gi/Go, the purified D2 receptors mediate the agonist stimulation of 35S-labeled guanosine 5'-O-(thiotriphosphate) binding to brain G-proteins with a typical D2-dopaminergic order of potency. These data suggest that we have purified an intact functional D2-dopamine receptor.

Affinity Labels↗