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Examination of Turnip Mosaic Virus Genome‐Linked Protein (VPg) as an Effective Novel Inhibitor of Ricin Toxin A Chain (RTA)
Author(s) -
De Los Santos Sabrina,
Domashevskiy Artem V
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.607.7
Subject(s) - ricin , ribosome inactivating protein , ribosome , biochemistry , rna , ribosomal rna , microbiology and biotechnology , ribosomal protein , biology , toxin , chemistry , gene
Ricin, a toxin originating from the seeds of Ricinus communis , is a type II ribosome inactivating protein (RIP) that is potent against eukaryotic ribosomes. The A chain of ricin, RTA, with functionality as an RNA N‐glycosidase, is capable of terminating protein synthesis via cleavage of the N‐glycosidic bond of a particular adenine residue in the highly conserved sarcin/ricin (S/R) loop of 28S ribosomal RNA. Pokeweed antiviral protein (PAP), stemming from the leaves of Phytolacca americana , is a type I RIP with both depurinating and antiviral properties. A previous study established that turnip mosaic virus genome‐linked protein (VPg), required for viral functions such as infectivity, inhibits the aforementioned characteristics of PAP. Based on the aforementioned study, we hypothesize that VPg will interact with RTA, relying on similarities in PAP and RTA active sites, and will inhibit RTA activity against ribosomal RNA. Following purification of VPg and RTA, steady state fluorescence will aid in the determination of the binding affinity pertaining to the protein‐protein complex. Quantification of the cleaved adenines, with and without VPg present, will be conducted through a bioluminescent assay that incorporates the conversion of adenine into adenosine triphosphate to produce luminescence via an enzymatic reaction involving firefly luciferase.