scholarly journals Characterization of Pokeweed Antiviral Protein (PAP) Isoforms and Comparison of Their Enzymatic Activities toward the Tobacco Etch Virus RNA

2015 ◽  
Vol 29 (S1) ◽  
Author(s):  
Valentina Aitbakieva ◽  
Artem Domashevskiy
Keyword(s):  
Tobacco Etch Virus ◽  
Enzymatic Activities ◽  
Pokeweed Antiviral Protein ◽  
Antiviral Protein ◽  
Virus Rna

scholarly journals Purification and partial characterization of another form of the antiviral protein from the seeds of Phytolacca americana L. (pokeweed)

1982 ◽  
Vol 203 (1) ◽  
pp. 55-59 ◽  
Author(s):  
L Barbieri ◽  
G M Aron ◽  
J D Irvin ◽  
F Stirpe
Keyword(s):  
Protein Synthesis ◽  
High Yield ◽  
Phytolacca Americana ◽  
Pokeweed Antiviral Protein ◽  
Antiviral Protein ◽  
Whole Cells ◽  
Reticulocyte Lysate ◽  
Simplex Virus

1. The pokeweed antiviral protein, previously identified in two forms (PAP and PAP II) in the leaves of Phytolacca americana (pokeweed) [Obrig. Irvin & Hardesty (1973) Arch. Biochem. Biophys. 155, 278-289; Irvin, Kelly & Robertus (1980) Arch. Biochem. Biophys. 200, 418-425] is a protein that prevents replication of several viruses and inactivates ribosomes, thus inhibiting protein synthesis. 2. PAP is present in several forms in the seeds of pokeweed. One of them, which we propose to call ‘pokeweed antiviral protein from seeds’ (PAP-S) was purified in high yield (180 mg per 100 g of seeds) by chromatography on CM-cellulose, has mol.wt. 30 000, and is similar to, but not identical with. PAP and PAP II. 3. PAP-S inhibits protein synthesis in a rabbit reticulocyte lysate with an ID50 (concentration giving 50% inhibition) of 1.1 ng/ml (3.6 × 10(-11) M), but has much less effect on protein synthesis by whole cells, with an ID50 of 1 mg/ml (3.3 × 10(-5) M), and inhibits replication of herpes simplex virus type 1.

Development of a double sandwich ELISA able to discriminate between free PAP (pokeweed antiviral protein) and complexed PAP in leaf extracts

Plant Science ◽  
1999 ◽  
Vol 140 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Jean-Luc Schlick ◽  
Bénédicte Desvoyes ◽  
Mehdi Hakil ◽  
Pascale Adami ◽  
Philippe Dulieu
Keyword(s):  
Sandwich Elisa ◽  
Pokeweed Antiviral Protein ◽  
Antiviral Protein ◽  
Leaf Extracts

scholarly journals Analysis of Ribonucleotide 5′-Triphosphate Analogs as Potential Inhibitors of Zika Virus RNA-Dependent RNA Polymerase by Using Nonradioactive Polymerase Assays

2016 ◽  
Vol 61 (3) ◽  
Author(s):  
Gaofei Lu ◽  
Gregory R. Bluemling ◽  
Paul Collop ◽  
Michael Hager ◽  
Damien Kuiper ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Zika Virus ◽  
Nonstructural Protein ◽  
Rna Dependent Rna Polymerase ◽  
Antiviral Compounds ◽  
Double Stranded Dna ◽  
Virus Rna ◽  
Potential Inhibitors

ABSTRACT Zika virus (ZIKV) is an emerging human pathogen that is spreading rapidly through the Americas and has been linked to the development of microcephaly and to a dramatically increased number of Guillain-Barré syndrome cases. Currently, no vaccine or therapeutic options for the prevention or treatment of ZIKV infections exist. In the study described in this report, we expressed, purified, and characterized full-length nonstructural protein 5 (NS5) and the NS5 polymerase domain (NS5pol) of ZIKV RNA-dependent RNA polymerase. Using purified NS5, we developed an in vitro nonradioactive primer extension assay employing a fluorescently labeled primer-template pair. Both purified NS5 and NS5pol can carry out in vitro RNA-dependent RNA synthesis in this assay. Our results show that Mn2+ is required for enzymatic activity, while Mg2+ is not. We found that ZIKV NS5 can utilize single-stranded DNA but not double-stranded DNA as a template or a primer to synthesize RNA. The assay was used to compare the efficiency of incorporation of analog 5′-triphosphates by the ZIKV polymerase and to calculate their discrimination versus that of natural ribonucleotide triphosphates (rNTPs). The 50% inhibitory concentrations for analog rNTPs were determined in an alternative nonradioactive coupled-enzyme assay. We determined that, in general, 2′-C-methyl- and 2′-C-ethynyl-substituted analog 5′-triphosphates were efficiently incorporated by the ZIKV polymerase and were also efficient chain terminators. Derivatives of these molecules may serve as potential antiviral compounds to be developed to combat ZIKV infection. This report provides the first characterization of ZIKV polymerase and demonstrates the utility of in vitro polymerase assays in the identification of potential ZIKV inhibitors.

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