Purification of 3 monomeric monocot mannose-binding lectins and their evaluation for antipoxviral activity: potential applications in multiple viral diseases caused by enveloped viruses

Three monomeric monocot lectins from Zephyranthes carinata, Zephyranthes candida, and Gloriosa superba with carbohydrate specificity towards mannose derivatives and (or) oligomannose have been isolated and purified from their storage tissues. The lectins were purified by anion-exchange chromatography on DEAE–Sephacyl and by gel filtration chromatography on Biogel P-200 followed by high-performance liquid chromatography. The purified lectins, Z. carinata, Z. candida, and G. superba had molecular masses of 12, 11.5, and 12.5 kDa, respectively, as determined by gel filtration and SDS–PAGE, indicating that they are monomers. In a hapten inhibition assay, methyl-α-d-mannopyranoside inhibited agglutination of both Z. candida and Z. carinata; the latter was also inhibited by Man(α1-2)Man and Man(α1-3)Man. Gloriosa superba showed inhibition only with Man(α1-4)Man of all of the sugars and glycoproteins tested. All purified lectins agglutinated red blood cells from rabbit, whereas G. superba was also reactive towards erythrocytes from guinea pig. All of the lectins were nonglycosylated and did not require metal ions for their activity. They were labile above 60 °C and were affected by denaturing agents such as urea, thiourea, and guanidine–HCl. The lectins were virtually nonmitogenic, like other members of Amaryllidaceae and Liliaceae. Of the 3 lectins, G. superba was found to be highly toxic to the BSC-1 cell line (African green monkey kidney epithelial cells), while both of the Zephyranthes species showed significant in vitro inhibition of poxvirus replication in BSC-1 cells without any toxic effects to the cells. In addition, Z. candida also exhibited significant anticancer activity against SNB-78, a CNS human cancer cell line.

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Brain peptides and glial growth. II. Identification of cells that secrete glia-promoting factors.

The Journal of Cell Biology ◽

10.1083/jcb.102.3.812 ◽

1986 ◽

Vol 102 (3) ◽

pp. 812-820 ◽

Cited By ~ 48

Author(s):

D Giulian ◽

D G Young

Keyword(s):

Nervous System ◽

High Performance ◽

Gel Filtration ◽

Anion Exchange Chromatography ◽

Brain Cell ◽

Exchange Chromatography ◽

Secretory Cells ◽

Brain Peptides ◽

The Central Nervous System

Glia-promoting factors (GPFs) are brain peptides which stimulate growth of specific macroglial populations in vitro. To identify the cellular sources of GPFs, we examined enriched brain cell cultures and cell lines derived from the nervous system for the production of growth factors. Ameboid microglia secreted astroglia-stimulating peptides, while growing neurons were the best source of the oligodendroglia-stimulating factors. These secretion products co-purified by gel filtration, anion exchange chromatography, and reverse-phase high performance liquid chromatography with GPFs isolated from goldfish and rat brain. Our findings suggest that glial growth in the central nervous system is regulated in part by a signaled release of peptides from specific secretory cells.

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New Amylase Inhibitor Present in Corn Seeds Active In Vitro Against Amylase from Fusarium verticillioides

Plant Disease ◽

10.1094/pdis.2003.87.3.233 ◽

2003 ◽

Vol 87 (3) ◽

pp. 233-240 ◽

Cited By ~ 8

Author(s):

Edson L. Z. Figueira ◽

Alejandro Blanco-Labra ◽

Antônio Carlos Gerage ◽

Elisabete Y. S. Ono ◽

Elizabeth Mendiola-Olaya ◽

...

Keyword(s):

High Performance ◽

Fusarium Verticillioides ◽

Gel Filtration ◽

Anion Exchange Chromatography ◽

Human Saliva ◽

Exchange Chromatography ◽

Experimental Conditions ◽

Amylase Inhibitor ◽

Fusarium Sp

A screening for specific amylase inhibitor levels against amylase from Fusarium verticillioides (Fusarium moniliforme), the most relevant mycotoxigenic fungus in corn, was conducted on 37 corn hybrids. The amylase inhibitor levels in these hybrids ranged from 5.5 to 16.0 amylase inhibitor units per gram of corn (AIU/g) in the MASTER and AG5011 hybrids, respectively. The hybrid with the maximum content of inhibitor was used as the source of this new protein. The inhibitor was partially purified using fractional precipitation, gel filtration on Sephadex G-75 column, high performance liquid chromatography (HPLC) Superose HR 10/30 column, and HPLC anion exchange chromatography, obtaining a 20.7-fold purification. Electrophoresis after denaturing and heating under reductive conditions showed an apparent 23.8 kDa molecular mass and an acidic isoelectric point of 5.4, which differs from previous molecular masses reported for other inhibitors present in corn seeds (14 and 22 kDa). This inhibitor showed activity against amylases from human saliva and pancreas, from the fungi F. verticillioides and Aspergillus flavus, and from the insects Acanthoscelides obtectus, Zabrotes subfasciatus, Tribolium castaneum, and Sitotroga cerealella. The mycoflora found in the corn grain indicated Fusarium sp. as the most prevalent fungi (81.1% of the samples), with a count ranging from 1.5 × 102 to 2.4 × 106 CFU/g of corn. The presence of fumonisins was detected in 21 out of the 37 hybrids studied, ranging from 0.05 to 2.67 μg of FB per gram of corn. No correlation could be established between this amylase inhibitor level in the corn seeds and the presence of Fusarium sp. or with the fumonisin content under the experimental conditions of the test.

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Co-purification of bone resorbing activity and adenylate cyclase stimulating activity from human tumours associated with the humoral hypercalcaemia of malignancy

Acta Endocrinologica ◽

10.1530/acta.0.1140018 ◽

1987 ◽

Vol 114 (1) ◽

pp. 18-26 ◽

Cited By ~ 5

Author(s):

Chohei Shigeno ◽

Itsuo Yamamoto ◽

Shegiharu Dokoh ◽

Megumu Hino ◽

Jun Aoki ◽

...

Keyword(s):

Bone Resorption ◽

High Performance ◽

Basic Protein ◽

Gel Filtration ◽

Exchange Chromatography ◽

Protein Factor ◽

Human Tumours ◽

Acid Stable ◽

Bone Resorbing Activity

Abstract. We have partially purified a tumour factor capable of stimulating both bone resorption in vitro and cAMP accumulation in osteoblastic ROS 17/2 cells from three human tumours associated with humoral hypercalcaemia of malignancy. Purification of tumour factor by sequential acid urea extraction, gel filtration and cation-exchange chromatography, reverse-phase high performance liquid chromatography followed by analytical isoelectric focussing provided a basic protein (pI > 9.3) with a molecular weight of approximately 13 000 as a major component of the final preparation which retained both the two bioactivities. Bone resorbing activity and cAMP-increasing activity in purified factor correlated with each other. cAMP-increasing activity of the factor was heat- and acid-stable, but sensitive to alkaline ambient pH. Treatment with trypsin destroyed cAMP-increasing activity of the factor. Synthetic parathyroid hormone (PTH) antagonist, human PTH-(3– 34) completely inhibited the cAMP-increasing activity of the factor. The results suggest that this protein factor, having its effects on both osteoclastic and osteoblastic functions, may be involved in development of enhanced bone resorption in some patients with humoral hypercalcaemia of malignancy.

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Evidence for N-linked glycosylation in Toxoplasma gondii

Biochemical Journal ◽

10.1042/bj2910713 ◽

1993 ◽

Vol 291 (3) ◽

pp. 713-721 ◽

Cited By ~ 29

Author(s):

M Odenthal-Schnittler ◽

S Tomavo ◽

D Becker ◽

J F Dubremetz ◽

R T Schwarz

Keyword(s):

Toxoplasma Gondii ◽

Gel Filtration ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Surface Glycoprotein ◽

Common Precursor ◽

The Common ◽

Oligosaccharide Structures

In this paper we report experiments demonstrating the presence of N-linked oligosaccharide structures in Toxoplasma gondii tachyzoites, providing the first direct biochemical evidence that this sporozoan parasite is capable of synthesizing N-linked glycans. The tachyzoite surface glycoprotein gp23 was metabolically labelled with [3H]glucosamine and [3H]mannose. Gel-filtration chromatography on Bio-Gel P4 columns produced four radiolabelled N-linked glycopeptides which were sensitive to peptidase-N-glycanase F, but resistant to endoglycosidases H and F. Using chemical analysis and exoglycosidase digestions followed by Dionex-high-pH anion-exchange chromatography and size fractionation on Bio-Gel P4 we show that gp23 has N-linked glycans in the hybrid- or complex-type structure composed of N-acetylgalactosamine, N-acetylglucosamine and mannose and devoid of sialic acid and fucose residues. In addition, the sensitivity of glycopeptides from glycoprotein extracts to endoglycosidases H and F revealed the in vivo synthesis of oligomannose-type structures by T. gondii tachyzoites. We have extended these findings by demonstrating the ability of T. gondii microsomes to synthesize in vitro a glucosylated lipid-bound high-mannose structure (Glc3Man9GlcNAc2) that is assumed to be identical with the common precursor for N-glycosylation in eukaryotes.

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A Novel Hemagglutinin with Antiproliferative Activity against Tumor Cells from the Hallucinogenic MushroomBoletus speciosus

BioMed Research International ◽

10.1155/2014/340467 ◽

2014 ◽

Vol 2014 ◽

pp. 1-7 ◽

Cited By ~ 7

Author(s):

Jian Sun ◽

Tzi-Bun Ng ◽

Hexiang Wang ◽

Guoqing Zhang

Keyword(s):

Cation Exchange ◽

Antiproliferative Activity ◽

Gel Filtration ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Lymphocytic Leukemia ◽

Hemagglutinating Activity ◽

Cation Exchange Chromatography ◽

Type Ab

Little was known about bioactive compounds from the hallucinogenic mushroomBoletus speciosus. In the present study, a hemagglutinin (BSH,B. speciosushemagglutinin) was isolated from its fruiting bodies and enzymatic properties were also tested. The chromatographic procedure utilized comprised anion exchange chromatography on Q-Sepharose, cation exchange chromatography on CM-Cellulose, cation exchange chromatography on SP-Sepharose, and gel filtration by FPLC on Superdex 75. The hemagglutinin was a homodimer which was estimated to be approximately 31 kDa in size. The activity of BSH was stable up to 60°C, while there was a precipitous drop in activity when the temperature was elevated to 70°C. BSH retained 25% hemagglutinating activity when exposed to 100 mM NaOH and 25 mM HCl. The activity was potently inhibited by 1.25 mM Hg2+and slightly inhibited by Fe2+, Ca2+, and Pb2+. None of the sugars tested showed inhibition towards BSH. Its hemagglutinating activity towards human erythrocytes type A, type B, and type AB was higher than type O. The hemagglutinin showed antiproliferative activity towards hepatoma Hep G2 cells and mouse lymphocytic leukemia cells (L1210)in vitro, with IC50of 4.7 μM and 7.0 μM, respectively. It also exhibited HIV-1 reverse transcriptase inhibitory activity with an IC50of 7.1 μM.

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Combination of high-performance anion-exchange chromatography and electrospray mass spectrometry for analysis of the in vitro O-glycosylated mucin motif peptide

Journal of Chromatography B Biomedical Sciences and Applications ◽

10.1016/0378-4347(94)00223-1 ◽

1994 ◽

Vol 658 (1) ◽

pp. 31-38 ◽

Cited By ~ 12

Author(s):

Daniel Tetaert ◽

Benoît Soudan ◽

Jean-Marc Lo-Guidice ◽

Colette Richet ◽

Pierre Degand ◽

...

Keyword(s):

Mass Spectrometry ◽

Anion Exchange ◽

High Performance ◽

Electrospray Mass Spectrometry ◽

Anion Exchange Chromatography ◽

Exchange Chromatography

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Suppressive Effect of Fig (Ficus Carica) Latex on Esophageal Cancer Cell Proliferation

Acta Facultatis Medicae Naissensis ◽

10.2478/v10283-012-0042-2 ◽

2013 ◽

Vol 30 (2) ◽

pp. 93-96 ◽

Cited By ~ 4

Author(s):

Seyyed Abbas Hashemi ◽

Saeid Abediankenari

Keyword(s):

Esophageal Cancer ◽

Cell Line ◽

Cancer Cell ◽

Human Cancer ◽

Cancer Cell Line ◽

Ficus Carica ◽

Esophageal Cancer Cell ◽

Fig Tree ◽

Vitro Effect

SUMMARY Fig (Ficus carica) tree latex was a source of treatment of different diseases in the Iranian traditional medicine reported by Avicenna in his 10th century book Canon of Medicine. The aim of this investigation was to establish the anticancer effect of fig tree latex on human cancer cells. The in vitro effect of different doses of fig tree latex including 2.5 mg/ml, 5 mg/ml, and 10 mg/ml on esophageal cancer cell line was evaluated after 72 hours by MTT assay. There was a significant change in 10 mg/ml treatment of latex after 72 hours on esophageal cancer line (P; 0.025). Ten mg/ml was the optimum concentration in the inhibition of cell line growth. Fig (Ficus carica) tree latex could be a candidate as a potential agent for the inhibition of cancerous cells production and development.

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Synthesis, Spectral Characterization, and In Vitro Cytotoxicity of Some Fe(III) Complexes Bearing Unsymmetrical Salen-Type Ligands Derived from 2-Hydroxynaphthaldehyde and Substituted Salicylaldehydes

Journal of Chemistry ◽

10.1155/2021/8028064 ◽

2021 ◽

Vol 2021 ◽

pp. 1-9

Author(s):

Quang Trung Nguyen ◽

Phuong Nam Pham Thi ◽

Nguyen Van Tuyen

Keyword(s):

Cell Line ◽

Cancer Cell ◽

Human Cancer ◽

Cancer Cell Line ◽

Tumor Cell Line ◽

In Vitro Cytotoxicity ◽

Ionization Mass ◽

Hek 293 ◽

Human Cancer Cell Lines

Six Fe(III) complexes bearing unsymmetrical salen-type ligands derived from 2-hydroxynaphthaldehyde and substituted salicylaldehydes were synthesized by coordinating the unsymmetrical salen-type ligands with FeCl3.6H2O. The synthetic complexes were characterized by electrospray ionization mass spectra (ESI-MS), effective magnetic moments (μeff), and infrared (IR) and ultraviolet-visible (UV-Vis) spectra. The spectroscopic data are in good agreement with the suggested molecular formulae of the complexes. Their cyclic voltammetric studies in acetonitrile solutions showed that the Fe(III)/Fe(II) reduction processes are electrochemically irreversible. The in vitro cytotoxicity of the obtained complexes was screened on human cancer cell lines KB (a subline of Hela tumor cell line) and HepG2 (a human liver cancer cell line) and a normal human cell line HEK-293 (Human Embryonic Kidney cell line). The results showed that the synthetic Fe(III) complexes are highly cytotoxic and quite selective. The synthetic complexes bearing unsymmetrical salen-type ligands with different substituted groups in the salicyl ring indicate different cytotoxicity.

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