Preterminal protein,the achilles heel of adenoviridae:Implications for adenoviral infections  

作　　者：Harold A Walsh 

机构地区：[1]Faculty of Pharmacy,Division of Pharmacology,Rhodes University,Grahamstown 6139,Eastern Cape,South Africa

出　　处：《World Journal of Pharmacology》2024年第2期1-9,共9页世界药理学杂志（英文）

摘　　要：BACKGROUND Adenoviruses pose a serious health risk particularly in the absence of any clinically approved treatment.As adenoviral infections are quite frequent and recent outbreaks manifest more virulent variant strains,the need to develop an effective treatment remains a priority.The adenoviral protein,preterminal protein(pTP),is one of the key common products of the viral lifecycle as it is necessary to initiate viral replication and hence the infection process.This makes pTP a potential chemotherapeutic target in the search for and development of an effective treatment for adenoviral induced infections.Here we report,for the first time,that glycosylation of pTP in situ prevents binding to ssDNA in vitro.AIM To explore whether specific structural tailoring of the adenoviral protein pTP,imparts the potential to scupper the viral replication process.METHODS All chemicals used were of reagent grade.Overexpression of pTP was achieved using the‘BAC to BAC’expression system.The presence and relative concentration of the protein was determined throughout the incubation period by the Bradford assay.The pTP was identified by MALDI-TOFF and sodium dodecyl sulphate polyacrylamide gel electrophoresis.For the removal of the aminosugar,a deglycosylase enzyme kit from PROZYME was used.Purification of cloned pTP(6xHis)was done with a ssDNA cellulose column followed by a Ni-NTA column.His-tags were excised with the Tobacco etch virus protease.Protein fractionation was performed with a fraction collector coupled to a UV detector(280 nm)from Pharmacia.RESULTS The pTP overexpressed in insect cells(Spodoptera frugiperda)(>96 hours),is unable to bind to ssDNA in vitro.Treatment of this unbound protein with a deglycosidase enzyme that is specific for the removal of truncated unsubstituted O-linked Galβ(1-3)GalNAc-α1 disaccharides bound to Thr or Ser in a glycoprotein,restores binding to ssDNA.Data is presented as a linegraph for both the glycosylated and the deglycosylated proteins.Each point represents the mean of triplicate e

关 键 词：Thomsen-Friedenreich antigen ADENOVIRAL Preterminal protein Spodoptera frugiperda cells Protein priming mechanism Glycosylation 

分 类 号：R965[医药卫生—药理学]