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1.
It has previously been shown in our laboratory that wheat germ agglutinin (WGA) binds to Trichoderma viride and inhibits growth of this fungus. Here we report on the effect of WGA, soybean agglutinin (SBA) and peanut agglutinin (PNA) on Penicillia and Aspergilli. Binding of the lectins to the fungi was examined with the aid of their fluorescein isothiocyanate (FITC) conjugated derivatives. FITC-WGA bound to young hyphal walls of all species, in particular to the hyphal tips and septa, in agreement with the chitinous composition of the cell walls of the two genera. Hyphae of all species examined were labelled, though in different patterns, by FITC-SBA and FITC-PNA, suggesting the presence of galactose residues on their surfaces. Young conidiophores, metulae (of the Penicillia), vesicles (of the Aspergilli), sterigmata and young spores, were also labelled. The three lectins inhibited incorporation of [3H]acetate, N-acetyl-D-[3H]glucosamine and D-[14C]galactose into young hyphae of Aspergillus ochraceus, indicating interference with fungal growth. Inhibition of spore germination by the three lectins was also observed. Preincubation of the lectins with their specific saccharide inhibitors prevented binding and the inhibitory effects. We conclude that lectins are useful tools for the study of fungal cell surfaces, and may also serve as an important aid in fungal classification. The present findings also support the suggestion that one role of lectins in plants is protection against fungal pathogens.Abbreviations Con A
concanavalin A
- PNA
peanut agglutinin
- SBA
soybean agglutinin
- WGA
wheat germ agglutinin
- FITC
fluorescein isothiocyanate
- GlcNAc
N-acetyl-D-glucosamine
- GalNAc
N-acetyl-D-galactosamine 相似文献
2.
Summary Sites of binding of eight different lectins (LTA, UEA I, WGA, SBA, DBA, CON A, PNA, RCA I) to cat submandibular gland were studied after exposure of tissue sections to sialidase, -fucosidase, -galactosidase, -mannosidase, -N-acetylglucosaminidase. All lectins were affected by enzymatic predigestion and the labeling of individual lectins was highly dependent upon the glycosidase used to pretreat the sections. Glycoconjugates of demilunar, acinar and ductal cells exhibited a different composition of terminal sequences. For example, fucose proved to form the disaccharide fucose-galactose in demilunar and acinar cells, whereas it was present with the sequence fucose-N-acetyl-d-glucosamine in striated duct cells. Sialic acid participated both to the terminal sequence sialic acid-galactose and sialic acid-N-acetyl-d-galactosamine either in demilunar or in ductal cells. Lectin labeling combined with glycosidase digestion was also helpful in verifying the influence of neighbouring oligosaccharides on the affinity of lectins for the respective sugars. 相似文献
3.
Summary Peroxidase-labelled lectins specific for various carbohydrate residues were used as histochemical reagents in the investigation of Hurler's syndrome. Peanut lectin was used to detect terminald-galactose, wheatgerm lectin forN-acetyl-d-glucosamine, soybean lectin forN-acetyl-d-galactosamine,Tetragonolobus lotus lectin for -l-fucose andBandeiraea S. lectin for -d-galactose. It was found that Kupffer cells in the liver and splenic reticulo-endothelial cells contain acid mucopolysaccharides which bind lectins in paraffin sections after appropriate fixation. The pattern of lectin binding suggests that such cells contain significant amounts ofd-galactose,l-fucose,N-acetyl-d-galactosamine andN-acetyl-d-glucosamine. It is likely that the last named carbohydrate is present as a polymer. Neurones contain a different carbohydrate, rich in galactose and fucose but poor inN-acetyl-d-glucosamine. This compound is resistant to lipid extraction. Hepatocytes, as a rule, do not react with lectins, most likely because of loss of the more soluble mucopolysaccharides during fixation. The results are consistent with the biochemical data of Hurler's syndrome and indicate that lectins can be a useful tool for the investigation of the cytochemistry of storage disorders. 相似文献
4.
S. Willemer H. Köhler R. Naumann H. F. Kern G. Adler 《Histochemistry and cell biology》1990,93(3):319-326
Summary Lectin-binding studies were performed at the ultrastructural level to characterize glycoconjugate patterns on membrane systems in pancreatic acinar cells of the rat. Five lectins reacting with different sugar moieties were applied to ultrathin frozen sections: concanavalin A (ConA): glucose, mannose; wheat-germ agglutinin (WGA): N-acetylglucosamine, sialic acid; Ricinus communis agglutinin I (RCA I): galactose; Ulex europaeus agglutinin I (UEA I): l-fucose; soybean agglutinin (SBA): N-acetylgalactosamine). Binding sites of lectins were visualized either by direct conjugation to colloidal gold or by the use of a three-step procedure involving additional immune reactions. The rough endoplasmic reticulum and the nuclear envelope of acinar cells was selectively labelled for ConA. The membranes of the Golgi apparatus bound all lectins applied with an increasing intensity proceeding from the cis-to the trans-Golgi area for SBA, UEA I and WGA. In contrast RCA I selectively labelled the trans-Golgi cisternae. The membranes of condensing vacuoles and zymogen granules were labelled for all lectins used although the density of the label differed between the lectins. In contrast the content of zymogen granules failed to bind SBA and WGA. Lysosomal bodies (membranes and content) revealed binding sites for all lectins used. The plasma membranes were heavily labelled by all lectins except for SBA which showed only a weak binding to the lateral and the apical plasma membrane. These results are in accordance to current biochemical knowledge of the successive steps in the glycosylation of membrane proteins. It could be demonstrated, that the cryo-section technique is suitable for the fine structural localisation of surface glycoconjugates of plasma membranes and internal membranes in pancreatic acinar cells using plant lectins. 相似文献
5.
Marilynn E. Etzler 《Glycoconjugate journal》1994,11(5):395-399
The carbohydrate binding properties of theDolichos biflorus seed lectin and DB58, a vegetative tissue lectin from this plant, were compared using two types of solid phase assays. Both lectins bind to hog blood group A + H substance covalently coupled to Sepharose 4B and this binding can be inhibited with free blood group A + H substance. However, the binding of the seed lectin is inhibited byD-GalNAc whereas DB58 binding was not inhbited by any monosaccharide tested, thus suggesting that its carbohydrate combining site may be more extensive than that of the seed lectin. The activities of these two lectins also differ from one another in ability to recognize blood group A + H substance adsorbed on to plastic and in the effects of salt and urea on their carbohydrate binding activities. Neither lectin showed glycosidase activity with p-nitrophenyl -D-GalNAc or p-nitrophenyl -D-GalNAc. 相似文献
6.
Summary Fluorescein or rhodamine conjugates of seventeen different lectins were tested for their ability to label the plasma membrane of live plant protoplasts. During the investigation, a strong effect of calcium was observed on the binding of several lectins to protoplasts derived from suspension cultured rose cells (Rosa sp. Paul's Scarlet). The binding of these lectins was increased by elevating the calcium concentration from 1 to 10 mM in the buffer. Other divalent cations had variable, but similar, effects on lectin binding. The mechanism of this effect appeared to involve the protoplast surface rather than the lectins. Although the cell wall-degrading enzymes used to isolate protoplasts had generally no effect on lectin binding, one clear exception was observed. Binding ofArachis hypogaea agglutinin was markedly reduced on protoplasts isolated with Driselase as compared to protoplasts isolated with a combination of Cellulysin and Pectolyase Y-23. Although most of the lectins that labeled protoplasts derived from cultured rose cells or from corn root cortex (Zea mays L. WF9 × Mo17) had specificities for galactose or N-acetylgalactosamine, some differences in protoplast labeling between lectins of the same saccharide specificity were observed. Two different analyses of the interaction betweenRicinus communis agglutinin and rose protoplasts showed that binding was cooperative with an apparent association constant of 7.2 × 105M–1 or 9.8 × 105M–1 with a maximum of approximately 108 lectin molecules bound per protoplast. Treatment of protoplasts with glycosidases which hydrolyze either N- or O-glycosidic linkages of glycoproteins slightly enhanced labeling of protoplasts byRicinus communis agglutinin. Interpretation of these results are discussed.Abbreviations MPR
medium, minimal organic medium (Nothnagel andLyon 1986)
- APA
Abrus precatorius agglutinin
- CSA
Cytisus sessilifolius agglutinin
- ECA
Erythrina cristagalli agglutinin
- GS-I
Griffonia simplicifolia agglutinin
- LcH
Lens culinarus agglutinin
- PNA
Arachis hypogaea agglutinin
- SBA
Glycine max agglutinin
- VAA
Viscum album agglutinin
- VFA
Vicia faba agglutinin
- WGA
Triticum vulgaris agglutinin
- Con A
Canavalia ensiformis agglutinin
- HPA
Helix pomatia agglutinin
- TPA
Tetragonolobus purpureas agglutinin
- RCA
Ricinus communis agglutinin
- DBA
Dolichos biflorus agglutinin
- SJA
Sophora japonica agglutinin
- BPA
Bauhinia purpurea agglutinin
- FITC
fluorescein isothiocyanate
- Ga1NAc
N-acetylgalactosamine
- FDA
fluorescein diacetate
- 2-O-Me-D-Fuc
2-O-methyl-D-fucose
Parts of the work presented here are also submitted in partial fulfillment of requirements for the Ph.D. degree. 相似文献
7.
Lokesh Bhattacharyya Martin Haraldsson Nathan Sharon Halina Lis Fred Brewer 《Glycoconjugate journal》1989,6(1):141-150
Erythrina lectins possess similar structural and carbohydrate binding properties. Recently, tri- and tetra-antennary complex type carbohydrates with non-reducing terminal galactose residues have been shown to be precipitated as tri- and tetravalent ligands, respectively, with certainErythrina lectins [Bhattacharyya L, Haraldsson M, Brewer CF (1988) Biochemistry 271034-41]. The present work describes a comparative study of the binding and precipitating activities of fourErythrina lectins,viz. E. corallodendron, E. cristagalli, E. flabelliformis, andE. indica, with multi-antennary complex type carbohydrates and synthetic cluster glycosides. The results show that though their binding affinities are very similar, theErythrina lectins show large differences in their precipitating activities with the carbohydrates. The results also indicate significant dependence of the precipitating activities of the lectins on the core structure of the carbohydrates. These findings provide a new dimension to the structure-activity relationship of the lectins and their interactions with asparagine-linked carbohydrates.Abbreviations EAL, ECorL, ECL, EFL, and EIL
represent the lectins from the seeds ofErythrina arborescens,
-
E. corallodendron, E. cristagalli, E. flabelliformis, andE. indica
respectively
- AFOS
thetri-antennary complex type oligosaccharide from asialofetuin
- AFGP
the tri-antennary glycopeptide from asialofetuin
- MeGal
methyl -d-galactopyranoside
Unless stated otherwise all sugars are in thed-configuration. 相似文献
8.
Lectins were extracted from whole fern grindings ofAzolla pinnata (AP) andAzolla filiculoides (AF) by precipitation with ammonium sulfate to 20% of saturation. At high pH both lectins dissociate into inactive subunits (5000 mol wt) which reassociate into active aggregates (>500,000 mol wt) following concentration by ammonium sulfate precipitation or freezing and thawing. Although amino sugars inhibited hemagglutinating activity of both AP and AF lectins,d-fructose was inhibitory only to the AP lectin hemagglutinating activity, andd-galactose was slightly inhibitory to the AP lectin but not to the AF lectin. Both lectins exhibited specificity for freshly extracted cyanobionts from homologous fern species: AP lectin agglutinated cyanobiont filaments from AP, but not from AF; AF lectin agglutinated cyanobiont filaments from AF, but not AP. Neither lectin reacted with cultured cyanobionts from either fern species. Hemagglutinating titers were likewise reduced by adsorption of these lectins to homologous cyanobiont cells. This report provides strong suggestive evidence for specificity in this N-fixing symbiosis between aquatic fern and cyanobacterium. 相似文献
9.
The subpopulations were compared of neurons in human dorsal root ganglia (DRG), as substance P, identified by somatostatin, Glycine max lectin (SBA) specific to terminal N-acetylgalactosamine, and Ulex europaeus I agglutinin (UEA-I) specific to l-fucose. The lectins and neuropeptides all bound to neurons of small diameter. Furthermore, the majority of the SBA binding neurons or somatostatin positive neurons were also UEA-I binding neurons. However, SBA binding neurons were not colocalized with somatostatin or substance P. Less than 20% of substance P positive neurons showed colocalization with l-fucosyl residues, and approximately 10% of l-fucosyl residues showed colocalization with substance P. Our results suggest that both l-fucose and terminal N-acetylgalactosamine containing neurons in the human DRG are subjected to different subpopulations from substance P or somatostatin positive neurons. 相似文献
10.
M I Khan M Joginadha Swamy M V Krishna Sastry S Umadevi Sajjan S R Patanjali Prasad Rao G V Swarnalatha P Banerjee A Surolia 《Glycoconjugate journal》1988,5(1):75-84
Fluorescence and stopped-flow spectrophotometric studies on three plant lectins fromPsophocarpus tetragonolobus (winged bean),Glycine max (soybean) andArtocarpus integrifolia (jack fruit) have been studied usingN-dansylgalactosamine as a fluorescent ligand. The best monosaccharide for the winged bean agglutinin I (WBA I) and soybean (SBA) is Me-GalNAc and for jack fruit agglutinin (JFA) is Me-Gal. Examination of the percentage enhancement and association constants (1.51×106, 6.56×106 and 4.17×105 M–1 for SBA, WBA I and JFA, respectively) suggests that the combining regions of the lectins SBA and WBA I are apolar whereas that of JFA is polar. Thermodynamic parameters obtained for the binding of several monosaccharides to these lectins are enthalpically favourable. The binding of monosaccharides to these lectins suggests that the-OH groups at C-1, C-2, C-4 and C-6 in thed-galactose configuration are important loci for interaction with these lectins. An important finding is that the JFA binds specifically to Galß1-3GaINAc with much higher affinity than the other disaccharides which are structurally and topographically similar.The results of stopped-flow spectrometry on the binding ofN-dansylgalactosamine to these lectins are consistent with a bimolecular single step mechanism. The association rate constants (2.4×105, 1.3×104, and 11.7×105 M–1 sec–1 for SBA, WBA I and JFA, respectively) obtained are several orders of magnitude slower than the ones expected for diffusion controlled reactions. The dissociation rate constants (0.2, 3.2×10–2, 83.3 sec–1 for SBA, WBA I and JFA, respectively) obtained for the dissociation ofN-dansylgalactosamine from its lectin complex are slowest for SBA and WBA I when compared with any other lectin-ligand dissociation process.Abbreviations SBA
Soybean agglutinin
- WBA I
Winged bean agglutinin (Basic)
- JFA
Jack fruit agglutinin
- PNA
Peanut agglutinin
- Con A
Concanavalin A
- Dansyl (Dns)
5-dimethylaminonaphthalene-I-sulphonyl
- 2GaINDns
N-dansylgalactosamine
- dGal
2-deoxygalactose
-
l-Ara
l-arabinose
-
d-Fuc
d-fucose
-
l-Rha
l-rhamnose
-
N-acetyllactosamine
Galß4GlcNAc
- melibiose
Gal6Glc 相似文献
11.
This work studied the effect of two cell-surface lectins isolated from the nitrogen-fixing soil bacterium Azospirillum brasilense Sp7 and from its mutant defective in hemagglutinating activity, A. brasilense Sp7.2.3, on the activities of α-glucosidase, β-glucosidase and β-galactosidase in the exocomponent, membrane and apoplast
fractions of wheat-seedling roots. Lectin (40 μg mL−1) incubation for 1 h of the plant fractions increased the enzymes’ activities; both wild-type and mutant lectins were most
stimulatory to the activities of all the exocomponent-fraction enzymes studied and to the apoplast-fraction β-glucosidase.
Pretreatment of the lectins with their carbohydrate hapten, L-fucose, lowered the effect. The observed differences in the lectins’ ability to influence enzyme catalytic activity are explained
by change in the antigenic properties of the mutant lectin. 相似文献
12.
Extracts from 44 species of Vietnamese marine algae, including 15 Chlorophyta, 18 Rhodophyta and 11 Phaeophyta species, were
examined for hemagglutination activity with a variety of different animal and human erythrocytes that were untreated or treated
with enzymes. Almost all extracts showed activity toward at least one type of erythrocytes, although those from three Chlorophyta
and two Rhodophyta species showed no hemagglutination with any type of erythrocytes examined. Strong activity was detected
in extracts from two Chlorophyta (Anadyomene plicata and Avrainvillea erecta) and four Rhodophyta species (Gracilaria eucheumatoides, Gracilaria salicornia, Kappaphycus alvarezii, and Kappaphycus striatum) with enzyme-treated rabbit and sheep erythrocytes. The hemagglutinins of seven Chlorophyta and eight Rhodophyta species
were examined for sugar-binding specificity, pH- and temperature-stability, and divalent cation-independency of hemagglutination
using ammonium sulfate-precipitates prepared from their extracts. In a hemagglutination-inhibition test with various monosaccharides
and glycoproteins, none of the hemagglutinins had affinity for monosaccharides, except the Codium arabicum and Gracilaria euchematoides hemagglutinins, whose activities were inhibited by both N-acetyl-d-galactosamine and N-acetyl-d-glucosamine. On the other hand, all of the hemagglutinins activities were inhibited by some glycoproteins. The inhibition
profiles with glycoproteins were different depending on hemagglutinin species, and suggest the presence of lectins specific
for high mannose N-glycans, complex N-glycans, or O-glycans. The activities of these algal hemagglutinins were stable over
a wide range of pH and temperature, and independent of the presence of divalent cations. These results indicate that Vietnamese
marine algae are a good source of novel and useful lectins. 相似文献
13.
Potato lectin (Solanum tuberosum agglutinin, STA) is an unusual glycoprotein containing approximately 50% carbohydrates by weight. Of the total carbohydrates,
92% is contributed by L-arabinose, which are O-linked to hydroxyproline residues. The ferric chloride-orcinol assay (Bial’s test), which is specific for pentoses has so
far been used only for the determination of free pentoses in biological samples. However, this colorimetric assay has not
been used for the detection of pentoses in bound form as it occurs in Solanaceae lectins (potato, tomato, and Datura lectins).
Utilizing the pentose colorimetric assay for monitoring the presence of potato lectin, a simpler and shorter procedure for
the purification of this lectin from potato tubers has been developed. The yield of potato lectin (1.73 mg per 100 g potato
tuber) is twice compared to the yields reported in earlier procedures. Although potato lectin is well known for its specificity
to free trimers and tetramers of N-acetyl-D-glucosamine (GlcNAc), it possesses a similar specificity to the core (GlcNAc)2 of N-linked glycoproteins. The utilization of the pentose assay in the purification of arabinose-rich lectins/agglutinins
obviates the necessity for the use of agglutination assay in the various purification steps. The pentose assay appears to
be a simple and convenient colorimetric assay for detecting any pentose-rich glycoprotein in plant extracts. The utility of
the pentose assay appears to have a significant potential in the detection of hydroxyproline-rich glycoproteins (HRGPs), which
are generally O-arabinosylated. 相似文献
14.
Summary Hand sections of young corn root tips have been used in a study of problems encountered in the binding of fluorescently-labelled lectins to plant tissue. It was found, surprisingly, that with lectins specific for a sugar known to be present (Lotus andUlex lectins forl-fucose), with a lectin specific for a sugar thought not to be present (wheat-germ agglutinin for N-acetylglucosamine), with non-lectin glycoprotein and protein (-globulin and bovine serum albumin) and with basophilic dyes (alcian blue and toluidine blue), a coincidental binding pattern similar to the pattern of autofluorescence in the same tissue was obtained. Corn root tissues include cell walls composed of complex polysaccharides esterified with ferulic acid residues, as well as mucilages which are highly hydrated and expanded. In such material, neither standard inhibition controls with haptens nor the use of a wide range of lectin concentrations are adequate to distinguish clearly specific and non-specific binding of fluorescently-labelled lectin. Therefore, lectins are not the simple test probes they have been supposed. Before interpreting results obtained in using fluorescently-labelled lectins on any tissue sections, all available information (biochemical as well as histochemical) about the tissue must be considered. 相似文献
15.
A. Bardosi L. Bardosi R. Lindenblatt H.-J. Gabius 《Histochemistry and cell biology》1990,94(3):285-291
Summary Different carrier-immobilized carbohydrate moieties were employed as tools to detect respective binding sites glycohistochemically and glycobiochemically. Besides ascertaining their presence the pattern of endogenous sugar receptors (lectins) in different regions of the human central nervous system was mapped to reveal any non-uniform expression. A strong and specific staining with biotinylated neoglycoproteins, exposing different sugar moieties as ligands, indicated the presence of sugar receptors in the nuclei, neuronal pathways and accessory structures such as ependyma cells, plexus chorioideus, intra- and extracerebral vessels and leptomeninges localized in the mesencephalon, in the pons, in the medulla oblongata and in the cerebellum. Significant differences were seen for various neuroanatomical regions like nerve cells in the basal and central regions of the nuclei pontis in the glycohistochemically detected level of expression of endogenous sugar receptors (lectins). The used approach with carbohydrate constituents of cellular glycoconjugates as ligands in search of specific receptors complemented studies on the localization of glycoconjugates with sugar-specific tools like plant lectins. Exemplary glycobiochemical investigations on the medulla oblongata and cerebellum were performed to investigate the molecular nature of sugar receptors detected glycohistochemically. Despite notable overall similarities, carbohydrate-binding proteins of differing molecular weight can be isolated from these two regions of the central nervous system, namely in the case of receptors with specificity to -galactoside termini, to N-acetyl-d-galactosamine and N-acetyl-d-glucosamine and to d-xylose. These combined glycohistochemical and glycobiochemical results serve as a guideline for exploring the physiological relevance of the detected regional differences. 相似文献
16.
Sgambati E Marini M Vichi D Zappoli Thyrion GD Parretti E Mello G Gheri G 《Histochemistry and cell biology》2007,128(3):263-273
The aim of this study was to investigate the distribution of the oligosaccharides of the glycoconjugates in placentas from
pregnancies complicated by different degree of altered glycaemia. Placentas from women with physiological pregnancies (group
1), with pregnancies complicated by minor degree of glucose intolerance (group 2) and with pregnancies complicated by gestational
diabetes mellitus (GDM) treated with insulin (group 3) were collected. Ten lectins were used (ConA, WGA, PNA, SBA, DBA, LTA,
UEA I, GSL II, MAL II and SNA) in combination with chemical and enzymatic treatments. The data showed a decrease of sialic
acid linked α(2–6) to galactose/N-acetyl-d-galactosamine and an increase of N-acetyl-d-glucosamine in the placentas of the pathological groups, in particular the group 3, comparing to the group 1. A decrease
of l-fucose (LTA) and d-galactose-(β1–3)-N-acetyl-d-galactosamine, and an increase and/or appearance of l-fucose (UEA I) and N-acetyl-d-galactosamine were observed in both the pathological groups, particularly in the group 2, with respect to the group 1. In
GDM, and even in pregnancies with a simple alteration of maternal glycaemia, the changes in the distribution of oligosaccharides
could be related to alteration of the structure and functionality of the placenta. 相似文献
17.
C. Meban 《The Histochemical journal》1986,18(4):196-202
Summary The surface coating of the pneumonocytes in human neonatal lung was studied by means of an electron microscope technique. Slices of aldehyde-fixed lung tissue were labelled with a horseradish peroxidase conjugate of one of the following lectins:Dolichos biflorus lectin,Triticum vulgaris lectin,Canavalia ensiformis lectin (concanavalin A),Limulus polyphemus lectin,Lotus tetragonolobus lectin andArachis hypogaea lectin. The tissue slices were then incubated in a diaminobenzidine—hydrogen peroxide medium and then postfixed in an osmium tetroxide solution. It was found that the type I and type II pneumonocytes were strongly labelled with the lectins ofTriticum vulgaris, Canavalia ensiformis, Limulus polyphemus andArachis hypogaea. The type I pneumonocytes were also strongly labelled withDolichos biflorus lectin but the staining of type II cells was relatively weak with this agent. Neither type of epithelial cell was labelled withLotus tetragonolobus lectin conjugate. These results suggest that the surface coating of the pneumonocytes in human neonatal lung contains the following carbohydrate groups:N-acetylgalactosamine,N-acetylglucosamine,-d-mannose,-d-galactose and sialic acid. 相似文献
18.
In wild-type Drosophila hydei (genotype X/Y) four different primary spermatocyte nuclear glycoproteins, classified as non-Y encoded because of their occurrence in X/O genotypes, were demonstrated to possess a few epitopes that depended on formation of the Y chromosomal giant lampbrush loops threads (th; Mr 55000 proteins) or pseudonucleolus (ps; Mr 38000, 58000 and 98000 proteins). The epitopes reacted with lectins and/or antibodies in vitro (lectin-/immunoreplica of primary spermatocyte total nuclear protein), and were lacking in mutants not possessing the respective loops. Those dependent on ps reacted with human sera. Epitopes restricted to proteins from th-forming spermatocytes reacted with lectin Con A (specific for d-Man and/or d-Glc) and antibodies directed against mouse immunoglobulins (AIA). In situ experiments (immunofluorescence microscopy of primary spermatocyte nuclei) revealed antibody cross-reactions with the respective loops. The reagents stained the distal (fused) sections and proximal (compact) parts of ps (human sera) or the proximal (compact) parts of th (AIA). Reaction with the latter loops was significantly repressed after absorption of AIA with the l-Fuc carbohydrate unit, classifying the AIA as fucosyl specific, and the epitopes along th as l-Fuc carbohydrate units.Abbreviations
D-man
D-mannose
-
D-Glc
D-glucose
-
L-Fuc
L-fucose
-
D-GlcNac
N-acetyl-D-glucosamine
-
D-GalNac
N-acetyl-D-galactosamine
-
AIA
anti-immunoglobulin antibody
-
hnRNP
heterogeneous nuclear ribonucleoprotein
-
DNP
deoxyribonucleoprotein
-
FITC
fluorescein isothiocyanate 相似文献
19.
An ultrastructural study of the morphology and lectin-binding properties of human mast cell granules
Summary The morphological characteristics and lectin-binding properties of mast cell granules from four human neurofibromata are described. Ultrastructural examination of the granules revealed that some contained dense cores, others had membranous configurations and some forms were intermediate between the two. A round electron-lucent area was present in some granules.After treatment with biotinylated lectins (10 g ml–1) followed by an avidin-peroxidase revealing system (5 g ml–1 in 0.125m Tris-buffered saline with 0.347m NaCl, pH 7.6), mast cell granules strongly bound Concanavalin A, garden pea, lentil, wheatgerm, erythro- and leuco-kidney bean lectins. This indicated the presence of abundantN-linked complex-type saccharide sequences. Soybean and peanut lectins showed only weak binding, while the presence of sparse -l-fucosyl terminals was indicated by the weak binding of winged pea lectin. The staining intensity of wheatgerm lectin was considerably reduced when incubated in the presence of its specific competing sugar tri-N-acetylchitotriose.Despite a wide variety of morphological differences between granules, all showed similar staining patterns and all granules within a single cell shared the same binding characteristics. 相似文献
20.
The polyphenol oxidase (PPO) of field bean (Dolichos lablab) is a tetramer made up of two subunits of mass 29,000 and 31,000 Da. The amino acid sequence of the tryptic peptides showed
approximately 90% sequence identity to the d-galactose specific legume lectins. The haemagglutinating activity of a pure and homogenous preparation of PPO measured using
human erythrocytes was 1261 HAU mg−1 protein and was inhibited by d-galactose. Purification by galactose-sepharose chromatography also indicated that the PPO and haemagglutinating activities
were associated with a single protein. Crude extracts of other legumes did not exhibit PPO activity, yet cross reacted with
anti-PPO antibodies. This dual function protein with PPO and haemagglutinating activity is unique to field bean. The two activities
are independent of each other occurring at different loci on the protein. These observations further evidence and strengthen
the assumption that galactose specific legume lectins have enzymatic function. Both PPO and lectins are proteins that play
a vital role in the defense mechanism of plants. The complementarity of these two simultaneous and independent powerful defense
mechanisms exhibited by a single protein renders it a candidate gene for the development of inbuilt plant protection. 相似文献