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

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
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.

scholarly journals Chemical identity of tryptensin with angiotensin

1980 ◽  
Vol 187 (3) ◽  
pp. 647-653 ◽  
Author(s):  
K Arakawa ◽  
M Yuki ◽  
M Ikeda
Keyword(s):  
Amino Acid ◽  
Thin Layer ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Human Plasma Protein ◽  
Plasma Protein Fraction ◽  
Layer Chromatography

Tryptensin, a vasopressor substance generated from human plasma protein fraction IV-4 by trypsin, has been isolated and the amino acid composition analysed. The procedures used for the isolation were: (a) adsorption of the formed tryptensin on Dowex 50W (X2; NH4+ form); (b) gel filtration through Sephadex G-25; (c) cation-exchange chromatography on CM-cellulose; (d) anion-exchange chromatography on DEAE-cellulose; (e) re-chromatography on CM-cellulose; (f) gel filtration on Bio-Gel P-2; (g) partition chromatography on high-pressure liquid chromatography. The homogeneity of the isolated tryptensin was confirmed by thin-layer chromatography and thin-layer electrophoresis. The amino acid analysis of the hydrolysate suggested the following proportional composition: Asp, 1; Val, 1; Ile, 1; Tyr, 1; Phe, 1; His, 1; Arg, 1; Pro, 1. This composition is identical with that of human angiotensin.

scholarly journals Evidence for N-linked glycosylation in Toxoplasma gondii

1993 ◽  
Vol 291 (3) ◽  
pp. 713-721 ◽  
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.

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

1986 ◽  
Vol 102 (3) ◽  
pp. 812-820 ◽  
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.

Production of recombinant IGF1 and its action on neuroblastoma cells in vitro

2018 ◽  
Vol 18 (4) ◽  
pp. 34-41
Author(s):  
Sergey A. Ishuk ◽  
Elena G. Bogomolova ◽  
Olga A. Dobrovolskaya ◽  
Alyona O. Akhmetshina ◽  
Daria S. Krasnoshchek ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Cation Exchange ◽  
Expression System ◽  
Neuroblastoma Cells ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Recombinant Insulin ◽  
Prokaryotic Expression System

This study aimed to develop a method for producing human recombinant insulin-like growth factor (IGF-1) based on a prokaryotic expression system and to characterize the highly purified protein. To achieve the study’s goal, the following methods were conducted: we performed automated chemical synthesis of DNA, constructed the expression plasmid, obtained Escherichia coli cell-producers of human recombinant IGF-1, cultivated the obtained producer cells with the induction of recombinant protein synthesis by isopropyl-β-D-1-thiogalactopyranoside and lactose, and purified human recombinant IGF-1 with affinity and cation exchange chromatography. The recombinant protein IGF-1 forms inclusion bodies during synthesis in Escherichia coli BL21 cells that contain plasmid pET28-IGF-1. Purified recombinant protein was obtained with a purity of 98% using affinity and cation exchange chromatography methods. The protein yield was 6 mg of human recombinant IGF-1 from 1 g of raw biomass. The resulting protein has the ability to protect Neuro 2a neuroblastoma cells from death caused by the deprivation of serum in the culture medium and can stimulate the differentiation of cells into neurons. Thus, a highly purified human recombinant IGF-1 was obtained. This protein has biological activity and is suitable for preclinical studies.

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

2007 ◽  
Vol 85 (1) ◽  
pp. 88-95 ◽  
Author(s):  
Amandeep Kaur ◽  
Sukhdev Singh Kamboj ◽  
Jatinder Singh ◽  
Rajinder Singh ◽  
Melissa Abrahams ◽  
...  
Keyword(s):  
Cell Line ◽  
High Performance ◽  
Human Cancer ◽  
Gel Filtration ◽  
Cancer Cell Line ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Gloriosa Superba ◽  
African Green Monkey Kidney

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.

scholarly journals Peroxidase and peroxidase-oxidase activities of isolated human myeloperoxidases

1987 ◽  
Vol 242 (3) ◽  
pp. 673-680 ◽  
Author(s):  
B E Svensson ◽  
K Domeij ◽  
S Lindvall ◽  
G Rydell
Keyword(s):  
Enzyme Activity ◽  
Cation Exchange ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Relative Molecular Mass ◽  
Cation Exchange Chromatography ◽  
Healthy Donors ◽  
Activity Measurements ◽  
Molecular Masses ◽  
Sites Of Action

Isolated neutrophils from healthy donors were used for the isolation of four highly purified forms of myeloperoxidase as determined by spectral (A430/A280 ratio 0.80-0.87) and enzyme-activity measurements. Although the myeloperoxidases exhibited different elution profiles on cation-exchange chromatography, gel filtration indicated similar relative molecular masses. When these forms were assayed for peroxidase and peroxidase-oxidase activities with several substrates, they all exhibited virtually the same specific activities. These results suggest that possible functional differences between the enzymes may be related to differences in their sites of action rather than to differences in enzyme activity. Myeloperoxidase from a patient with chronic myeloid leukaemia also revealed a similar heterogeneity on cation-exchange chromatography. However, this myeloperoxidase contained in addition one form with a lower and one form with a higher relative molecular mass, as indicated by gel-filtration chromatography.

Isolation and Partial Characterization of Adipotropin, an Apparent New Lipolytic Peptide from Bovine Anterior Pituitary Lobes

1980 ◽  
Vol 35 (9) ◽  
pp. 1171-1177 ◽  
Author(s):  
Karen O’Malley ◽  
Karl Folkers ◽  
Olav Trygstad ◽  
Irene Foss
Keyword(s):  
Anterior Pituitary ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Frozen Tissue ◽  
Pituitary Glands ◽  
Approximate Molecular Weight ◽  
One Year

A peptide, apparently new, has been isolated from the anterior lobes of bovine pituitary glands by the following steps: (1) lyophilization of frozen tissue; (2) homogenization; (3) extraction with an aqueous buffer; (4) gel filtration; (5) anion and then (6) cation exchange chromatography; (7) HPLC. One antioxidant and two proteolytic inhibitors were present during buffer extraction to avoid artifactual reactions. Amino acid analyses consistently revealed the same amino acids and no others on different specimens, one year apart. The composition was estimated as: 4 Asp, 2 Thr, 5 Ser, 8 Glu(x), 5 Pro, 10 Gly, 4 Ala, 2 Val, 2 Met, 1 Ile, 2 Leu, 1 Tyr, 1 Phe, 2 His, 3 Lys, 2 Arg (5600 daltons). Cysteine and tryptophan were absent. An approximate molecular weight by gel filtration was 7200 daltons. The isoelectric point was 6.1. The peptide showed a dose-response activity in the range of 0.001-1.0 μg in a rabbit adipose system, in vitro. The peptide is tentatively designated as adipotropin, because it has no unequivocal chemical relation­ship to other relevant lipolytic peptides, and because it has the highest molar potency, in vitro, of these peptides, including pACTH.

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

Plant Disease ◽  
2003 ◽  
Vol 87 (3) ◽  
pp. 233-240 ◽  
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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