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141.
142.
Mistletoe toxic lectins consist of two polypeptide chains: an enzymatically active A chain, which is a toxic component, and a disulfide-bonded B chain, which confers the lectin properties on the total molecule. Mistletoe leaves contain three toxic lectins encoded by three genes. The B chains of these lectins were overproduced in Escherichia coli in a soluble form. The recombinant proteins bound with asialofetuin, but had substantially lower affinity for simple sugars D-galactose and N-acetyl-D-galactosamine as compared with the natural proteins. The functional properties of the B chains strongly depended on the storage conditions (salt concentration and the presence of galactose); the dependence was explained by structural instability of nonglycosylated recombinant proteins. The lectin activity of one of the recombinant B chains was close to that of the native protein, which was attributed to the lack of N-glycosylation sites in the latter. 相似文献
143.
A plant lectin was isolated from barley (Hordeum vulgare) coleoptiles using acidic extraction and different chromatographic methods. Sequencing of more than 50% of the protein sequence
by Edman degradation confirmed a full-length cDNA clone. The subsequently identified open reading frame encodes for a 15 kDa
protein which could be found in the soluble fraction of barley coleoptiles. This protein exhibited specificity towards mannose
sugar and is therefore, accordingly named as Horcolin (Hordeum vulgare coleoptile lectin). Database searches performed with
the Horcolin protein sequence revealed a sequence and structure homology to the lectin family of jacalin-related lectins.
Together with its affinity towards mannose, Horcolin is now identified as a new member of the mannose specific subgroup of
jacalin-related lectins in monocot species. Horcolin shares a high amino acid homology to the highly light-inducible protein
HL#2 and, in addition to two methyl jasmonic acid-inducible proteins of 32.6 and 32.7 kDa where the jasmonic acid-inducible
proteins are examples of bitopic chimerolectins containing a dirigent and jacalin-related domain. Immunoblot analysis with
a cross-reactive anti-HL#2 antibody in combination with Northern blot analysis of the Horcolin cDNA revealed tissue specific
expression of Horcolin in the coleoptiles. The function of Horcolin is discussed in the context of its particular expression
in coleoptiles and is then compared to other lectins, which apparently share a related response to biotic or abiotic stress
factors. 相似文献
144.
145.
Rakesh Mohan Kestwal Emadeldin Hassan E. Konozy Chwan-Deng Hsiao Maria Cristina Roque-Barreira Shobhana V. Bhide 《Biochimica et Biophysica Acta (BBA)/General Subjects》2007
α-mannosidase from Erythrina indica seeds is a Zn2+ dependent glycoprotein with 8.6% carbohydrate. The enzyme has a temperature optimum of 50 °C and energy of activation calculated from Arrhenius plot was found to be 23 kJ mol− 1. N-terminal sequence up to five amino acid residues was found to be DTQEN (Asp, Thr, Gln, Glu, and Asn). In chemical modification studies treatment of the enzyme with NBS led to total loss of enzyme activity and modification of a single tryptophan residue led to inactivation. Fluorescence studies over a pH range of 3–8 have shown tryptophan residue to be in highly hydrophobic environment and pH change did not bring about any appreciable change in its environment. Far-UV CD spectrum indicated predominance of α-helical structure in the enzyme. α-Mannosidase from E indica exhibits immunological identity with α-mannosidase from Canavalia ensiformis but not with the same enzyme from Glycine max and Cicer arietinum. Incubation of E. indica seed lectin with α-mannosidase resulted in 35% increase in its activity, while no such activation was observed for acid phosphatase from E. indica. Lectin induced activation of α-mannosidase could be completely abolished in presence of lactose, a sugar specific for lectin. 相似文献
146.
一种从食用真菌Agrocybe aegerita中提取的凝集素AAL具有显著的抗肿瘤效果。该蛋白的基因序列已经通过RT-PCR得到。T7噬菌体展示库筛选发现该蛋白可以在体外和细胞核内的细胞周期调控蛋白mortality factor related gene on chro-mosome 15(MRG15)发生相互作用。本实验目的是证实AAL和MRG15在细胞核内的共定位,为AAL与MRG15的体内的互作提供证据。构建质粒,将aal与mrg15分别连接到pEGFP-C1和pDsRed-C1上,共转染Hela细胞。将pEGFP-C1和pD-sRed-C1空载体共转染Hela细胞作为对照。利用激光共聚焦显微镜研究AAL和MRG15在细胞内的共定位。AAL和MRG15均定位在细胞核内,荧光图片的重叠表明AAL和MRG15在核内共定位。而对照组中的EGFP和DS-RED在细胞内呈弥散分布。这些为AAL和MRG15在细胞核内的相互作用提供了证据。 相似文献
147.
Maria Lígia Rodrigues Macedo Maria das Graas Machado Freire Maria Barbosa Reis da Silva Luana Cassandra Breitenbach Barroso Coelho 《Comparative biochemistry and physiology. Part A, Molecular & integrative physiology》2007,146(4):486
Bruchid beetle larvae cause major losses in grain legume crops throughout the world. Some bruchid species, such as the cowpea weevil (Callosobruchus maculatus) and the Mexican bean weevil (Zabrotes subfasciatus), are pests that damage stored seeds. The Mediterranean flour moth (Anagasta kuehniella) is of major economic importance as a flour and grain feeder; it is often a severe pest in flour mills. Plant lectins have been implicated as antibiosis factors against insects. Bauhinia monandra leaf lectin (BmoLL) was tested for anti-insect activity against C. maculatus, Z. subfasciatus and A. kuehniella larvae. BmoLL produced ca. 50% mortality to Z. subfaciatus and C. maculatus when incorporated into an artificial diet at a level of 0.5% and 0.3% (w/w), respectively. BmooLL up to 1% did not significantly decrease the survival of A. kuehniella larvae, but produced a decrease of 40% in weight. Affinity chromatography showed that BmoLL bound to midgut proteins of the insect C. maculatus. 33 kDa subunit BmoLL was not digested by midgut preparations of these bruchids. BmoLL-fed C. maculatus larvae increased the digestion of potato starch by 25% compared with the control. The transformation of the genes coding for this lectin could be useful in the development of insect resistance in important agricultural crops. 相似文献
148.
Ponpimol Tipthara Polkit Sangvanich Marcus Macth Amorn Petsom 《Journal of Plant Biology》2007,50(2):167-173
A mannose-binding lectin was isolated from rhizomes of the medicinal plantCurcuma zedoaria. We used extraction with 20 mM phosphate buffer, ammonium sulfate precipitation, ion exchange chromatography on Q-Sepharose,
gel filtration chromatography on Superdex 75, and reverse-phase HPLC. The purified lectin yielded a single band on SDS-PAGE
that corresponded to a molecular mass of 13 kDa. This lectin exhibited hemagglutinating activity toward rabbit erythrocytes,
which could be inhibited by mannose only. The lectin was digested with trypsin and its digests were analyzed using MALDI-TOF/TOF.
Partial amino acid sequences were obtained from tandem mass spectra via automatedde novo sequencing, and were then identified by MS-BLAST homology searches to enable recognition of related proteins in other species.
Inferred peptide sequences exhibited similarity to a mannose-binding lectin fromEpipactis helleborine, a member of the Orchidaceae. 相似文献
149.
An increasing number of galectins have been found in various animal species, the most abundant of which is galectin-1. The purpose of the present study was to purify and characterize galectin-1 from buffalo brain. We purified the galectin using a combination of ammonium sulphate fractionation and affinity chromatography and the homogeneity was determined by both native polyacrylamide gel electrophoresis (PAGE) and denaturing SDS-PAGE. The molecular weight of the galectin as determined by SDS-PAGE under reducing conditions and by gel filtration column under native conditions was 13.8 and 24.5 kDa, respectively, suggesting a dimeric form of galectin. The most potent inhibitor of the galectin activity was lactose, giving complete inhibition of hemagglutination at 0.8 mM. Galectin showed higher specificity towards human blood group A. Free thiol groups were estimated at a molar ratio of 2.9. The effects of alkylating reagents (iodoacetate and iodoacetamide) on saccharide binding of the galectin were studied. Both alkylating reagents significantly inactivated the activity of the galectin within 20 min. The temperature and pH stability of the galectin were determined. Our findings based on physico-chemical properties, carbohydrate and blood group specificities of the galectin may have future implications in biological and clinical applications. 相似文献
150.
Florian Wagner Karsten R Heidtke Bernd Drescher Uwe Radelof 《Briefings in Functional Genomics and Prot》2007,6(3):163-170
A number of fundamental technical developments like the evolvement of oligonucleotide microarrays, new sequencing technologies and gene synthesis have considerably changed the character of genomic biological resource centres in recent years. While genomic biological resource centres traditionally served mainly as providers of sparsely characterized cDNA clones and clone sets, there is nowadays a clear tendency towards well-characterized, high-quality clones. In addition, major new service units like microarray services have developed, which are completely independent of clone collections, reflecting the co-evolution of data generation and technology development. The new technologies require an increasingly higher degree of specialization, data integration and quality standards. Altogether, these developments result in spin-offs of highly specialized biotech companies, some of which will take a prominent position in translational medicine. 相似文献