scholarly journals Concanavalin A-agglutinability of membrane-skeleton-free vesicles and aged cellular remnants derived from human erythrocytes. Is the membrane skeleton required for agglutination?

1987 ◽  
Vol 241 (2) ◽  
pp. 513-520 ◽  
Author(s):  
S M Gokhale ◽  
N G Mehta
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Membrane Skeleton ◽  
Con A ◽  
Intact Cells ◽  
Rigid Skeleton

Vesicles and cell remnants have been obtained by aging of erythrocytes in vitro. The vesicles lacking the membrane skeletal proteins and the remnants known to possess a rigid skeleton have been used to assess the role of membrane skeletal proteins in the process of Con A (concanavalin A)-mediated agglutination of erythrocytes. Both the vesicles and the remnants were found to bind Con A at the same density as did intact cells. The vesicles, isolated from normal as well as from the Con A-agglutinable trypsin- and Pronase-treated cells, failed to agglutinate with Con A. They were, however, well agglutinated by WGA (wheat-germ agglutinin) and RCA [Ricinus communis (castor bean) agglutinin], indicating that the vesicles are not defective in agglutination. Large, cytoskeleton-free, vesicles prepared by another procedure also gave the same results. The aged remnants from trypsin- and Pronase-treated erythrocytes showed significantly decreased agglutination with Con A, but were agglutinated as well as the fresh cells by WGA and RCA. The agglutination with Con A is thus abolished when the membrane skeleton is absent, and reduced when it is rigid, suggesting that the skeleton may play an important role in the agglutination of erythrocytes by Con A.

Changing surface antigen and carbohydrate patterns during the development of Oesophagostomum dentatum

Parasitology ◽  
1999 ◽  
Vol 119 (5) ◽  
pp. 491-501 ◽  
Author(s):  
A. JOACHIM ◽  
B. RUTTKOWSKI ◽  
A. DAUGSCHIES
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Soybean Agglutinin ◽  
Peanut Agglutinin ◽  
Oesophagostomum Dentatum ◽  
Periodate Treatment

Living and fixed specimen of Oesophagostomum dentatum were labelled in situ with serum antibodies or a panel of biotin- labelled lectins. Specific binding of antibodies was observed in all parasitic stages – freshly exsheathed 3rd-stage larvae (L3), 3rd- and 4th-stage (L4) larvae cultured in vitro and L3 and L4 and adults isolated from pig intestines. The shedding of the stained layer by motile larvae was inhibited by levamisole-induced paralysis. Larvae cultured in vitro exposed serum-derived proteins on their surface which could be labelled with secondary antibody directed against the respective serum donor species. While freshly exsheathed larvae were recognized by O. dentatum-positive serum only, older larvae and adults cross-reacted with serum from pigs infected with O. quadrispinulatum, a closely related species. Lectin binding varied considerably between stages. While binding was not observed in pre-parasitic stages, Concanavalin A, Soybean Agglutinin, Wheat Germ Agglutinin, Ricinus communis Agglutinin and Peanut Agglutinin bound to developing larvae in varying degrees. Dolichos biflorus Agglutinin only bound to advanced (luminal) larval stages, while adults generally displayed only weak or partial lectin binding (except with Concanavalin A and Wheat Germ Agglutinin). Ulex europaeus Agglutinin only labelled larvae derived from cultures containing 10% pig serum. Cleavage of the carbohydrate residues by sodium periodate treatment resulted in reduction of antibody binding to cultured larvae, but not to freshly exsheathed L3. Concanavalin A, Soybean Agglutinin, and Peanut Agglutinin binding was also reduced by periodate treatment, while binding of Wheat Germ Agglutinin and Ricinus communis Agglutinin was inhibited only in early L3, but not in older stages. The different lectin labelling patterns are related to the different stages of the nematode – infective, invasive, histotropic, and luminal – and may serve as a mode of adaptation for the parasite against the host's immune attack by surface glycoprotein variation, together with antigen shedding (as demonstrated by labelling of motile larvae) and a possible acquisition of host molecules at the parasite's surface. Furthermore, a possible role of this developmental variation in surface carbohydrates in parasite–parasite interactions is discussed.

Lack of binding of serum glycoprotein hormone α subunit to Concanavalin A–Sepharose reflects increased branching of the oligosaccharide chains

1984 ◽  
Vol 103 (1) ◽  
pp. 111-116 ◽  
Author(s):  
A. J. Chapman ◽  
J. T. Gallagher ◽  
C. G. Beardwell ◽  
S. M. Shalet
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Lectin Binding ◽  
Pituitary Tumours ◽  
Con A ◽  
Glycoprotein Hormone ◽  
Α Subunit ◽  
Binding Forms

ABSTRACT The lectin-binding properties of serum α subunit were studied by lectin affinity chromatography. Normal individuals and most patients with pituitary tumours produced α subunit which bound specifically to Concanavalin A–Sepharose (Con A). Some patients with pituitary tumours produced both Con A-reactive α subunit and α subunit which did not bind to Con A. Concanavalin A–Sepharose-binding α subunit from all sources bound strongly to Ricinus communis agglutinin–Sepharose after treatment with neuraminidase. Serum α subunit from those patients with pituitary tumours, which did not bind to Con A, bound to wheat germ agglutinin–Sepharose, exhibiting both weakly binding and strongly binding forms. Serum α subunit from both patients and controls, which did bind to Con A, showed only weak affinity for wheat germ agglutinin–Sepharose. Neither the low affinity nor the high affinity of serum α subunit from any source for wheat germ agglutinin–Sepharose was affected by neuraminidase. These findings show that (a) the predominant pattern of glycosylation of serum α subunit from normal controls is a Con A-reactive, biantennate complex oligosaccharide and (b) that the structural alteration which results in serum α subunit which does not bind to Con A in some patients with pituitary tumours is not an absence of carbohydrate, rather the α subunit contains highly branched, either complex or hybrid oligosaccharides. J. Endocr. (1984) 103, 111–116

Lectin-mediated agglutination of amphibian embryonic cells

1977 ◽  
Vol 27 (1) ◽  
pp. 227-243
Author(s):  
B.R. Fraser ◽  
S.E. Zalik
Keyword(s):  
Concanavalin A ◽  
Wheat Germ ◽  
Cytochalasin B ◽  
Ricinus Communis ◽  
Soya Bean ◽  
Embryonic Cells ◽  
Glutaraldehyde Fixation ◽  
Con A ◽  
Neuraminidase Treatment ◽  
Ricinus Communis Agglutinin

Dissociated blastula cells of Xenopus laevis are agglutinated with wheat germ agglutinin (WGA), Ricinus communis agglutinin (RCA), concanavalin A (Con A) and, to a lesser extent with soya bean agglutinin (SBA). They are not agglutinated with fucose-binding protein. Neuraminidase treatment of cells enhances their agglutinability with RCA and SBA, but has no effect on Con A- and WGA-mediated agglutinability. Treatment of cells with procaine, or xylocaine, has no effect on the cells' agglutinability or on the extrusion of lobopodia. Treatment with colchicine or cytochalasin B either separately or simultaneously has no effect on lectin-mediated agglutinability. Cells treated with cytochalasin B or colchicine and cytochalasin B simultaneously lack lobopodial extensions, while colchicine alone has no effect on these structures. Phenothiazine tranquillizers inhibit agglutination mediated by all of the above mentioned lectins. Lobopodial extensions are absent in cells treated with these compounds. Glutaraldehyde fixation inhibits RCA and WGA mediated agglutinability and reduces the Con A-mediated agglutinability. Results suggest that in this system microtubules and microfilaments are not involved in lectin-mediated agglutination.

scholarly journals Ultrastructural localization of concanavalin A and wheat germ agglutinin binding sites in adult and embryonic mouse myocardium.

1982 ◽  
Vol 30 (3) ◽  
pp. 193-200 ◽  
Author(s):  
D Gros ◽  
B Bruce ◽  
C E Challice ◽  
J Schrevel
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ultrastructural Localization ◽  
Embryonic Cells ◽  
Con A ◽  
Embryonic Mouse ◽  
Transverse Tubular System ◽  
Receptor Sites ◽  
Ultrathin Sections

The lectins, concanavalin A (Con A) and wheat germ agglutinin (WGA), have been used to localize with precision glycosyl residues in adult and embryonic mouse myocardium. They were detected by means of an affinity method using peroxidase and chitobiosylperoxidase, respectively, which then were revealed with 3,3'-diaminobenzidine and H2O2. Exhaustive controls have shown that the binding of Con A and WGA is reversible when experiments are performed with adult specimens (tissue blocks or ultrathin sections of glycol methacrylate-embedded material) or with isolated embryonic cells. Experiments carried out with tissue blocks from embryonic hearts have shown peroxidase binding. This finding is discussed on the basis of the presence of the endogenous lectin-like components in embryonic hearts. Results show that the surface of adult and embryonic myocardial cells specifically bind both Con A and WGA, thus indicating the presence of glycosyl residues similar to alpha-methyl-D-mannoside and N-acetyl-D-glucosamine. In adult heart the transverse tubular system was also labeled. The absence of Con A and WGA receptor sites in the gap junction regions was demonstrated by means of an electron microscope postembedding staining method.

Evidence for heterogeneity of glycosylation of human renin obtained by using lectins

1991 ◽  
Vol 81 (3) ◽  
pp. 393-399 ◽  
Author(s):  
Masayuki Hosoi ◽  
Shokei Kim ◽  
Kenjiro Yamamoto
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Positive Staining ◽  
Carbohydrate Structure ◽  
Renal Secretion ◽  
Human Renin ◽  
Lentil Lectin ◽  
Ricinus Communis Agglutinin

1. In this study, the carbohydrate structure of pure human renin was examined by using various lectins. 2. Pure renin could be separated into three forms by concanavalin A chromatography, a concanavalin A-unbound form, a loosely bound form and a tightly bound form, termed renins A, B and C, respectively. Renins A, B and C accounted for 3, 13 and 84%, respectively, of the purified renin. These forms were all present in individual human plasma and the relative proportions in plasma were 27 ± 3, 33 ± 4 and 39 ± 5% (means ± sem) for renins A, B and C, respectively (n = 5). 3. Each form, electroblotted on to the nitrocellulose sheet after gel electrophoresis, was incubated with five peroxidase-labelled lectins, lentil lectin, erythroagglutinating phytohaemagglutinin, wheat-germ agglutinin, Ricinus communis agglutinin and peanut agglutinin. The protein was stained with 4-chloro-l-naphthol. 4. The staining pattern obtained with these lectins was significantly different among the three forms of human renin, confirming that they have different carbohydrate structures. Furthermore, the positive staining of human renin with erythroagglutinating phytohaemagglutinin, wheat-germ agglutinin and Ricinus communis agglutinin was in contrast with the lack of binding of rat renin to these lectins. 5. These results indicate the renal secretion of differently glycosylated multiple forms of human renin. The carbohydrate structure of human renin appears to differ from that of rat renin.

scholarly journals Freeze-fracture cytochemistry: partition of glycophorin in freeze-fractured human erythrocyte membranes.

1982 ◽  
Vol 93 (2) ◽  
pp. 463-469 ◽  
Author(s):  
P P da Silva ◽  
M R Torrisi
Keyword(s):  
Human Erythrocyte ◽  
Binding Sites ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Transmembrane Protein ◽  
Freeze Fracture ◽  
Membrane Skeleton ◽  
Erythrocyte Membranes ◽  
Con A ◽  
Human Erythrocyte Membranes

Thin-section and critical-point-dried fracture-labeled preparations are used to determine the distribution and partition of glycophorin-associated wheat germ agglutinin (WGA) binding sites over protoplasmic and exoplasmic faces of freeze-fractured human erythrocyte membranes. Most wheat germ agglutinin binding sites are found over exoplasmic faces. Label is sparse over the protoplasmic faces. These results contrast with previous observations of the partition of band 3 component where biochemical analysis and fracture-label of concanavalin A (Con A) binding sites show preferential partition of this transmembrane protein with the protoplasmic face. Presence of characteristic proportions of WGA and Con A binding sites over each fracture face is interpreted to indicate the operation of a stochastic process during freeze-fracture. This process appears modulated by the relative expression of each transmembrane protein at either surface as well as by their association to components of the erythrocyte membrane skeleton.

scholarly journals STUDI HISTOKIMIA LEKTIN TERHADAP JENIS DAN DISTRIBUSI GLIKOKONJUGAT ABOMASUM KERBAU RAWA (Bubalus bubalis) KALIMANTAN SELATAN

2015 ◽  
Vol 9 (2) ◽  
Author(s):  
Anni Nurliani ◽  
Teguh Budi Pitojo ◽  
Dwi Liliek Kusindarta
Keyword(s):  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Periodic Acid ◽  
Bubalus Bubalis ◽  
Soybean Agglutinin ◽  
Con A ◽  
Periodic Acid Schiff ◽  
Ulex Europaeus ◽  
Ricinus Communis Agglutinin

Penelitian ini bertujuan mengkaji efisiensi pencernaan kerbau rawa dengan mengidentifikasi jenis dan distribusi glikokonjugat  pada daerah abomasum kerbau rawa. Enam ekor kerbau rawa jantan >2,5 tahun dan berat badan 300-400 kg digunakan dalam penelitian ini. Sampel diperoleh dari rumah potong hewan (RPH) Kabupaten Banjar, Kalimantan Selatan. Setiap bagian abomasum meliputi kardiak, fundus, dan pilorus diambil untuk pengamatan mikroskopis dengan pewarnaan hematoksilin-eosin (HE) dan alcian blue-periodic acid schiff (AB-PAS). Residu gula glikokonjugat pada abomasum dideteksi dengan pewarnaan histokimia lektin dengan menggunakan wheat germ agglutinin (WGA), ricinus communis agglutinin (RCA), concanavalin agglutinin (Con A), ulex europaeus agglutinin (UEA), dan soybean agglutinin (SBA). Hasil penelitian menunjukkan bahwa daerah kardiak mengandung glikokonjugat D manosa/D glukosa, D galaktosa, dan N asetilglukosamin.  Daerah fundus mengandung D manosa/D glukosa, D galaktosa, L fukosa, N asetilglukosamin, dan N asetilgalaktosamin. Daerah pilorus mengandung glikokonjugat L fukosa dan N asetilglukosamin. Pola reaktivitas daerah kardiak, fundus, dan pilorus kerbau rawa terhadap pewarnaan histokimia lektin memiliki pola yang berbeda dengan ruminansia lain. Jenis glikokonjugat yang dimiliki oleh kerbau rawa tersebut diduga berkaitan dengan fungsi peningkatan kemampuan efisiensi pencernaan kerbau rawa. Setiap bagian abomasum kerbau rawa memiliki jenis glikokonjugat yang spesifik dengan pola distribusi khas sesuai dengan fungsinya.

Glycoconjugates in the human fetal endolymphatic sac as detected by lectins

1991 ◽  
Vol 105 (9) ◽  
pp. 711-715 ◽  
Author(s):  
Hiroshi Yamashita ◽  
Dan Bagger-Sjöbäck ◽  
Jan Wersäll ◽  
Toru Sekitani
Keyword(s):  
Epithelial Cell ◽  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Human Fetus ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Endolymphatic Sac ◽  
Maturation Process

AbstractThe distribution of glycoconjugates in the 11 to 16 weeks old human fetal endolymphatic sac (ES) was analyzed using six biotinylated lectins; Wheat germ agglutinin (WGA), Abrusprecatorius agglutinin (APA), Ulexeuropaeus agglutinin I (UEA-I), Ricinus communis agglutinin 120 (RCA120), Helixpomatia agglutinin (HPA), Concanavalin A (ConA). In the 11 week old human fetus, fluorescent reactions with WGA, APA, RCA120 and ConA were detected in the ES. There was almost no reaction with HPA and UEA-I. In the 14 week old human fetus, however.fluorescent reactions with HPA and UEA-I appeared. This result suggests that the presence of glycoconjugates changes during the maturation process of the ES. Glycoconjugates detected with HPA were related to the epithelial cell elements of the ES epithelium. The reaction with UEA-I suggests that the stainable substance present in the ES lumen may be secreted locally by the ES itself.

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