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1.
The direct double-antibody enzymelinked immunosorbent assay system was used in the detection and measurement of seed lectins from peanut (Arachis hypogaea L.) and soybean (Glycine max L.) plants (PSL and SBL, respectively) that had been inoculated with their respective rhizobia. Concentrations of PSL dropped to undetectable levels in peanut roots at 9 d and stems and leaves at 27 d after planting; SBL could no longer be detected in soybean roots at 9 d and in stems and leaves at 12 d. A lectin antigenically similar to PSL was first detected in root nodules of peanuts at 21 d reaching a maximum of 8 g/g at 29 d then decreasing to 2.5 g/g at 60 d. There was no evidence of a corresponding lectin in soybean nodules.Sugar haemagglutination inhibition tests with neuraminidase-treated human blood cells established that PSL and the peanut nodule lectin were both galactose/lactose-specific. Further tests with rabbit blood cells demonstrated a second mannosespecific lectin in peanut nodule extracts that was not detected in root extracts of four-week-old inoculated plants or six-week-old uninoculated plants, although six-week-old root extracts from inoculated plants showed weak lectin activity. The root extracts from both nodulated and uninoculated plants contained another peanut lectin that agglutinated rabbit but not human blood cells. Haemagglutination by this lectin was, however, not inhibited by simple sugars but a glycoprotein, asialothyroglobulin, was effective in this respect.Abbreviations DAS
double antibody sandwich
- ELISA
enzyme-linked immunosorbent assay
- PBS
phosphate-buffered saline
- PSL
peanut seed lectin
- SBL
soybean lectin 相似文献
2.
Cosme R. Martínez Alessandro V.P. Albertini Márcia V.B. Figueiredo Valdinete L. Silva Alexandre H. Sampaio Benildo S. Cavada José L. Lima-Filho 《World journal of microbiology & biotechnology》2004,20(1):77-83
High molecular weight lectins (> 100 kDa) from seeds of the legumes Canavalia brasiliensis (CnBr), Cratylia floribunda (CFL), Phaseolus vulgaris (PHA) and Vatairea macrocarpa (VML), temporarily stimulate the respiration of Rhizobium tropici-CIAT899 and R. etli-CFN42. These stimulants were significant (P < 0.05) in bacterial suspensions (> 2.85 mg dry biomass ml–1), having at least 6200 molecules of lectins per bacteria. The VML (20 g ml–1), induced specific O2 demand of 2.3–2.5 M O2 min–1 mg dry biomass–1, in CFN42 and CIAT899, respectively. However, CnBr, CFL and PHA induced smaller demands of O2 (5×), in both strains. The order of affinities of the lectins was approximately VML > PHA > CFL > CnBr, with regard to respiratory stimuli in CIAT899 strain. The co-administration of 10 g VML ml–1 and 9.8 M galactose, in CIAT899 suspensions, reduced the respiratory stimuli significantly in relation to the treatment with VML alone. These respiratory stimuli, induced by the lectins, increase the significance of the interaction lectin × Rhizobium in terms of bacterial physiology. Its understanding could be important in relation to bacterial symbiotic behaviour. 相似文献
3.
Molecular cloning of mannose-binding lectins from Clivia miniata 总被引:1,自引:0,他引:1
Els J. M. Van Damme Koen Smeets Fred Van Leuven Willy J. Peumans 《Plant molecular biology》1994,24(5):825-830
Screening of a cDNA library constructed from total RNA isolated from young developing ovaries of Clivia miniata Regel with the amaryllis lectin cDNA clone resulted in the isolation of four different isolectin clones which clearly differ from each other in their nucleotide sequences and hence also in their deduced amino acid sequences. Apparently the lectin is translated from an mRNA of ca. 800 nucleotides encoding a precursor polypeptide of 163 amino acids. Northern blot analysis of total RNA isolated from different tissues of Clivia miniata has shown that the lectin is expressed in most plant tissues with very high lectin mRNA concentrations in the ovary and the seed endosperm. 相似文献
4.
Pierre Rougé Carl A K Borrebaeck Michael Richardson Alan Yarwood 《Glycoconjugate journal》1987,4(4):371-378
The structure-function relationship of eight structurally very similarLathyrus lectins, which are two-chain lectins from theVicieae tribe, has been studied. Hemagglutination activity, inhibition of hemagglutination, glycoprotein reactivity and mitogenic activity were determined for each of the lectins. Despite the close structural resemblance, marked differences in the carbohydrate binding activity of theLathyrus lectins were recorded. These functional differences were related to small conformational changes due to minor differences in amino acid sequence of eachLathyrus lectin. 相似文献
5.
Subunit exchange between lectins from different cereal species 总被引:1,自引:0,他引:1
Lectins from Triticum monococcum, Secale cereale (rye), and Hordeum vulgare (barley) can exchange their subunits in vitro and thereby form (intergeneric) heteromeric lectins. An analysis of the isolectin pattern of a Triticale variety revealed that intergeneric heterodimers of wheat and rye lectin subunits are normal constituents of the embryo cells. It appears, therefore, that these different cereal lectins are structurally so closely related that their subunits can not distinguish between identical and nonidentical partners when they associate into dimers.Abbreviations CL
cereal lectin
- SP Sephadex
sulfopropyl Sephadex
- WGA
wheat germ agglutinin 相似文献
6.
Snowdrop and wheatgerm lectins and avidin as antimetabolites for the control of sugarcane whitegrubs 总被引:2,自引:0,他引:2
Snowdrop and wheatgerm lectins were found to be insecticidal and growth inhibiting dietary proteins for larvae of the sugarcane whitegrub Antitrogus parvulus. At concentrations as low as 0.5 mg of snowdrop lectin per gram of semi-artificial diet, growth was inhibited by 21 days of feeding and significant mortality was apparent by 28 days. Wheatgerm lectin was active at similar concentrations, although expression of the effects was slower. Avidin was found to be a growth inhibiting dietary protein for larvae of Antitrogus consanguineus. At levels as low as 0.01 mg g-1 of diet, growth was inhibited by 28 days of feeding. Avidin caused no significant mortality after 35 days of feeding. Snowdrop and wheatgerm lectins and avidin are insect growth-inhibiting proteins whose genes potentially could be manipulated into sugarcane and improve host-plant resistance to whitegrubs. 相似文献
7.
Gu Ji-Dong Belay Brook Mitchell Ralph 《World journal of microbiology & biotechnology》2001,17(2):173-179
A catheter surface was modified by coating a cellulose acetate polymer. Adhesion of Pseudomonas aeruginosa ATCC 27853 to the surface was investigated by exposing bacterial cultures to three treatments: polymer impregnated with silver ions (Ag+), polymer surfaces coated with lectins and a combination of Ag+ and a lectin coating. The effective concentration of Ag+ providing protection against bacterial biofilm development was 100g/ml and higher. Lectins alone at 10% also showed inhibition of bacterial attachment. However, the best result was achieved against bacterial adhesion and growth on surfaces using a combination of 100 g Ag+/ml and a lectin coating as a surface treatment. This surface treatment was also effective against both fresh culture and a two-week-old culture containing P. aeruginosa producing exopolymers. Our results suggest that Ag+impregnation combined with a lectin coating warrants further investigation as a potential means of protecting catheters. 相似文献
8.
Growth of most Rhizobium strains is inhibited by mimosine, a toxin found in large quantities in the seeds, foliage and roots of plants of the genera Leucaena and Mimosa. Some Leucaena-nodulating strains of Rhizobium can degrade mimosine (Mid+) and are less inhibited by mimosine in the growth medium than the mimosine-nondegrading (Mid-) strains. Ten Mid+ strains were identified that did not degrade 3-hydroxy-4-pyridone (HP), a toxic intermediate of mimosine degradation. However, mimosine was completely degraded by these strains and HP was not accumulated in the cells when these strains were grown in a medium containing mimosine as the sole source of carbon and nitrogen. The mimosine-degrading ability of rhizobia is not essential for nodulation of Leucaena species, but it provides growth advantages to Rhizobium strains that can utilize mimosine, and it suppresses the growth of other strains that are sensitive to this toxin. 相似文献
9.
The lectins from the seedsof Dolichos lablab var.lignosus (field bean) andDolichos lablab var.typicus (lablab bean) have been isolated in a homogeneous form by affinity chromatography on D-mannose linked Sepharose. Both the
lectins are glycoproteins and have a molecular weight of 60,000 andS
20,w value of 5.2 and seem to be made up of 4 similar subunits (apparent molecular weight 15,000). The carbohydrate content ofthe
lectins is mostly fucose (2–5 mol per mol of protein), mannose (5–8 mol per mol of protein) and N-acetyl glucosamine (1–2
mol per mol of protein). The amino acid composition of both the lectins was similar and methionine and half cystine could
not be detected, Both the lectins have similar tryptic peptide map. Alanine and serine were the only N and C-terminal amino
acids for both lectins. The lectins were found to contain low amounts of bound metals such as manganese, magnesium and calcium.
The near ultra-violet circular dichroism spectra of the lectins are similar to that of Sainfoin. Circular dichroism data indicate
that tyrosine and tryptophan residues are involved in sugar binding.
The lectins are nonspecific for human blood groups and they agglutinate a variety of other erythrocytes. Among a number of
sugars, D-glucose and D-mannose inhibited the haemag-glutinating activity ofthe lectins. The lectins were antigenically similar 相似文献
10.
The glucose-specific peanut root lectin, PRA II, is localized on the surface of 7-day-old peanut seedling root and in root cortical parenchymatous cells. The lectin is eluted from intact roots upon washing with buffer containing glucose. Rabbit erythrocytes bind to the root surface and the cortical cells; the binding is inhibited by antibodies raised against PRA II, peanut-specificRhizobium cells and by glucose. Lipopolysaccharides isolated from host-specificRhizobium strain inhibit the haemagglutinating activity of PRA II and are precipitated by the lectin. Our results suggest that PRA II might be involved in recognition ofRhizobium by peanut roots. 相似文献
11.
The seed lectins of black locust (robinia pseudoacacia) are encoded by two genes which differ from the bark lectin genes 总被引:2,自引:0,他引:2
Els J. M. Van Damme Annick Barre Pierre Rougé Fred Van Leuven Willy J. Peumans 《Plant molecular biology》1995,29(6):1197-1210
Two lectins were isolated from Robinia pseudoacacia (black locust) seeds using affinity chromatography on fetuin-agarose, and ion exchange chromatography on a Neobar CS column. The first lectin, R. pseudoacacia seed agglutinin I, referred to as RPsAI, is a homotetramer of four 34 kDa subunits whereas the second lectin, referred to as RPsAII, is composed of four 29 kDa polypeptides. cDNA clones encoding the polypeptides of RPsAI and RPsAII were isolated and their sequences were determined. Both polypeptides are translated from mRNAs of ca. 1.2 kb encoding a precursor carrying a signal peptide. Alignment of the deduced amino acid sequences of the different clones indicates that the 34 and 29 kDa seed lectin polypeptides show 95% sequence identity. In spite of this striking homology, the 29 kDa polypeptide has only one putative glycosylation site whereas the 34 kDa subunit has four of these sites. Carbohydrate analysis revealed that the 34 kDa possesses three carbohydrate chains whereas the 29 kDa polypeptide is only partially glycosylated at one site. A comparison of the deduced amino acid sequences of the two seed and three bark lectin polypeptides demonstrated unambiguously that they are encoded by different genes. This implies that five different genes are involved in the control of the expression of the lectins in black locust.Abbreviations LECRPAs
cDNA clone encoding Robinia pseudoacacia seed lectin
- LoLI
Lathyrus ochrus isolectin I
- PsA
Pisum sativum agglutinin
- RPbAI
Robinia pseudoacacia bark agglutinin I
- RPbAII
Robinia pseudoacacia bark agglutinin II
- RPsAI
Robinia pseudoacacia seed agglutinin I
- RPsAII
Robinia pseudoacacia seed agglutinin II 相似文献
12.
Xiulin Wang Liangyu Gong Shengkang Liang Xiurong Han Chenjian Zhu Yanbin Li 《Harmful algae》2005,4(2):6377
The algicidal activity of the rhamnolipid biosurfactants (the mixture of Rha-Rha-C10-C10 and Rha-C10-C10) produced by Pseudomonas aeruginosa was investigated in the present paper. The results indicated that the biosurfactants had potential algicidal effects on the harmful algal bloom (HAB) species, Heterosigma akashiwo. The growth of H. akashiwo was strongly inhibited in medium containing rhamnolipids (0.4–3.0 mg L−1); moreover, the rhamnolipids showed strong lytic activity toward H. akashiwo at higher concentrations (≥4.0 mg L−1). In addition, the effects of the rhamnolipids on the growth of Gymnodinium sp. and Prorocentrum dentatum, another two kinds of HAB species, were also studied. Compared with the dramatic algicidal effect on H. akashiwo, the cells of P. dentatum were inhibited or lysed at higher concentrations (1.0–10.0 mg L−1), while the cells of Gymnodinium sp. were not suppressed with the same treatment, indicating the rhamnolipids had the potential for the selective control of HABs.Morphometric analysis at ultrastructural level by transmission electron micrographs indicated that the extent of ultrastructural damage of the alga was severe at high concentrations of rhamnolipids and during extended periods of contact. The first response occurred in the plasma membrane which partly disintegrated. The lack of membrane facilitated the rhamnolipid biosurfactants into the cells and allowed damage to other organelles, which resulted in the injury of chloroplast, vacuolization of mitochondria and deformation of the cristae, disruption of nuclear membrane and condensation of chromatin in nucleus, suggesting that the lytic activity of rhamnolipids was mainly due to their powerful surfactivity and their tendency to cohere on the surface of phospholipids bimolecular layer of the cells and further destroyed the layers, and then the structure of quasi-membrane configurations inside the cells was disintegrated, following by the irreversible damage to the ultrastructure and the loss of the function of organelles, consequently leading the cells to lyse. 相似文献
13.
14.
Neoglycoconjugates are useful tools to study carbohydrate/protein interactions. In order to discover new lectins, to define their fine specificity or to study their intracellular trafficking, there is a need for neoglycoconjugates containing complex oligosaccharides. We recently set up a simple way to transform native oligosaccharides into glycosynthons. The present paper describes i[emsp4 ]) the synthesis of such glycosynthons starting with sialylated oligosides, ii[emsp4 ]) the preparation of sialylated neoglycoproteins and iii[emsp4 ]) their binding to sialic acid-specific lectins assessed by surface plasmon resonance experiments. 相似文献
15.
Two lectins were purified by affinity chromatography from mature peanut (Arachis hypogaea L.) nodules, and compared with the previously characterised seed lectin of this plant. One of the nodule lectins was similar to the seed lectin in its molecular weight and amino-acid composition and ability to bind derivatives of galactose. However, unlike the seed lectin, this nodule lectin appeared to be a glycoprotein and the two lectins were only partially identical in their reaction with antibodies prepared against the seed lectin. The other nodule lectin also appeared to be a glycoprotein but bound mannose/glucose-like sugar derivatives, and differed from the seed lectin in molecular weight, antigenic properties and amino-acid composition.Abbreviations Gal
galactose
- Gle
glucose
- GNL
galactose-binding nodule lectin
- Fru
fructose
- MNL
mannosebinding nodule lectin
-
M
r
rerative molecular mass
- PBS
phosphate-buffered saline
- PSL
peanut seed lectin
- SDS
sodium dodecyl sulphate
- Sorb
sorbitol 相似文献
16.
James E. Stewart 《Harmful algae》2011,10(6):706-712
Reduction or binding of Fe(III) by agent(s) produced by a highly toxigenic strain of Alexandrium excavatum was detected at approximately the same levels in culture filtrates of this dinoflagellate grown either axenically or non-axenically. Nanomolar concentrations of pure Paralytic Shellfish Poisons (PSP) produced by this phytoplankton, the carbamate toxins saxitoxin, 2/3 gonyautoxin or the N-sulfamoyl carbamate toxins C1/C2, added to a Schwyn and Neilands (1987) assay mixture did not stimulate Fe(III) reduction or binding. In contrast, additions of the N-sulfamoyl carbamate toxin, gonyautoxin V (GTXV also known as B1) alone resulted in a several-fold increase in this activity. The level of activity per cell was considerably higher for those cultures grown in media deficient in iron than in those whose growth was restricted by decreases of either N or P. 相似文献
17.
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
18.
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
19.
Dr. J. W. S. Brown T. C. Osborn F. A. Bliss Dr. T. C. Hall 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1982,62(4):361-367
Summary The relationship between the polypeptide composition and the agglutination behaviour of the lectin-containing G2/albumin protein groups has allowed the identification of the active lectin polypeptides in different cultivars of Phaseolus vulgaris (Brown et al. accompanying paper). These results were used to ascertain the particular G2/albumin group contained in the various lectin sources used previously for the purification of lectin proteins. Many studies were found to have included lectin sources which contained the same G2/albumin pattern (TG2) and this common denominator has permitted the direct comparison of the properties reported for these purified lectins. Thus, much of the extensive literature on bean lectins is concurred. 相似文献
20.
The location and topography of infection sites in soybean (Glycine max (L.) Merr.) root hairs spot-inoculated with Rhizobium japonicum have been studied at the ultrastructural level. Infections commonly developed at sites created when the induced deformation of an emerging root hair caused a portion of the root-hair cell wall to press against an adjacent epidermal cell, entrapping rhizobia within the pocket between the two host cells. Infections were initiated by bacteria which became embedded in the mucigel in the enclosed groove. Infection-thread formation in soybean appears to involve degradation of mucigel material and localized disruption of the outer layer of the folded hair cell wall by one or more entrapped rhizobia. Rhizobia at the site of penetration are separated from the host cytoplasm by the host plasmalemma and by a layer of wall material that appears similar or identical to the normal inner layer of the hair cell wall. Proliferation of the bacteria results in an irregular, wall-bound sac near the site of penetration. Tubular infection threads, bounded by wall material of the same appearance as that surrounding the sac, emerge from the sac to carry rhizobia roughly single-file into the hair cell. Growing regions of the infection sac or thread are surrounded by host cytoplasm with high concentrations of organelles associated with synthesis and deposition of membrane and cell-wall material. The threads follow a highly irregular path toward the base of the hair cell. Threads commonly run along the base of the hair cell for some distance, and may branch and penetrate into subjacent cortical cells at several points in a manner analagous to the initial penetration of the root hair. 相似文献