scholarly journals Postembedding staining of Brunner's gland with lectin-ferritin conjugates.

1981 ◽  
Vol 29 (8) ◽  
pp. 946-952 ◽  
Author(s):  
S Suzuki ◽  
S Tsuyama ◽  
T Suganuma ◽  
N Yamamoto ◽  
F Murata
Keyword(s):  
Concanavalin A ◽  
Wheat Germ Agglutinin ◽  
Present Method ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Staining Intensity ◽  
Ricinus Communis Agglutinin ◽  
Brunner’S Gland ◽  
Brunner's Gland ◽  
Embedding Methods

Postembedding staining of intracellular carbohydrates of rat Brunner's gland cells embedded in Epon and acrylamide was carried out with Ricinus communis agglutinin-ferritin, concanavalin A-ferritin, and wheat germ agglutinin-ferritin conjugates. Th Golgi vacuoles and mucous granules were stained with these conjugates. In each staining, the tissues embedded in acrylamide were stained more strongly than those embedded in Epon. The staining intensity of wheat germ agglutinin-ferritin was the strongest among the three conjugates and the staining intensity of Ricinus communis agglutinin-ferritin was stronger than that of concanavalin A-ferritin in both embedding methods. Free ferritin showed almost no binding to these structures and staining with the conjugates was inhibited by the addition of appropriate competitive sugars to the staining solutions. Osmium-postfixed tissues were not stained well with the conjugates. Washing of the sections with bovine serum albumin solution after staining was an essential step in the present method to reduce the nonspecific adsorption of the conjugates. The present method was very simple and had good reproducibility.

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.

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.

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 Wheat-germ-agglutinin and Ricinus communis-agglutinin-binding sites of BHK cells compared with each other and with 140 kDa fibronectin receptors

1988 ◽  
Vol 251 (1) ◽  
pp. 269-277 ◽  
Author(s):  
T L Tuan ◽  
F Grinnell
Keyword(s):  
Cell Surface ◽  
Binding Sites ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Ricinus Communis ◽  
Chinese Hamster ◽  
Surface Binding ◽  
Bhk Cells ◽  
Receptor Complexes ◽  
Ricinus Communis Agglutinin

We compared the wheat-germ agglutinin (WGA) and Ricinus communis agglutinin (RCA) binding sites of baby-hamster kidney (BHK) cells. There were 1.01 × 10(8) WGA-binding sites per cell (Kd = 0.027 nM) and 6 × 10(6) RCA-binding sites per cell (Kd = 0.014 nM). Binding of WGA or RCA to BHK cells resulted in more than 75% of the cell-surface binding sites becoming associated with the cytoskeleton (i.e. resistant to extraction with detergent), although no more than 10% of these sites were associated with the cytoskeleton before addition of the lectins. After binding of WGA to the cells, the cell surface was cross-linked so extensively that it remained intact even after detergent extraction of the treated cells, and could be observed by electron microscopy. A similar cross-linking effect did not occur after binding of RCA to cells, which may be because there were so many more binding sites for WGA than for RCA. The composition of WGA- and RCA-binding molecules was analysed by lectin affinity chromatography of metabolically radiolabelled BHK cells. We found that in the WGA-binding-molecule preparations there were eight major polypeptides, ranging in molecular mass from 93 to 340 kDa, and that the RCA-binding molecules were a subpopulation of the WGA-binding molecules. A polyclonal antibody against the 140 kDa fibronectin (FN) receptors of Chinese-hamster ovary (CHO) cells immunoblotted a 145 kDa polypeptide component in both WGA- and RCA-binding-molecule preparations. The results indicated that the 145 kDa component was present in at least two FN-receptor complexes that differed in glycosylation, only one of which was able to bind to RCA affinity columns. The oligomeric nature of the FN-receptor complex, which contained three polypeptides with molecular masses of 120-145 kDa, was demonstrated by using anti-(CHO-cell FN receptor) antibodies to immunoprecipitate extracts prepared from radioiodinated BHK cells.

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

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.

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.

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