Targeting enteroviral 2A protease by a 16-mer synthetic peptide: inhibition of 2Apro-induced apoptosis in a stable Tet-on HeLa cell line.
Enteroviridae such as coxsackievirus are important infectious agents causing viral heart diseases. Viral protease 2A (2Apro) initiates the virus life cycle, and is an excellent target for developing antiviral drugs. Here, to evaluate the validity of the 2Apro as a proper therapeutic target, and based on the existing information and molecular dynamics, a 16-mer peptide was designed to specifically target the active site of protease 2Apro in order to block the activity of CVB3 2Apro. We showed that the peptide could compete with endogenous substrate in a concentration-dependent manner. Further, we established a HeLa cell line that expressed 2Apro. Expression of 2Apro resulted in significant morphological alteration and eventual cell death. Western blot and viability assay showed that the 16-mer peptide (200 microg/ml) could significantly block 2Apro activity and its cytotoxic effect. Future modification of the 16-mer peptide can improve its affinity for 2Apro and therefore develop effective antiviral drug.
Maghsoudi
Nader
N
Neuroscience Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran. Electronic address: nmaghsoudi@sbmu.ac.ir.
Tafreshi
Narges Kh
NK
Neuroscience Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran; H. Lee Moffitt Cancer Center, Tampa, FL, USA.
Khodagholi
Fariba
F
Neuroscience Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran.
Zakeri
Zahra
Z
Department of Biology, Queens College and Graduate Center of the City University of New York, Flushing, NY, USA.
Esfandiarei
Mitra
M
Child & Family Research Institute, Department of Anesthesiology, Pharmacology & Therapeutics, University of British Columbia, Vancouver, Canada.
Hadi-Alijanvand
Hamid
H
Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran.
Sabbaghian
Marjan
M
Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran.
Maghsoudi
Amir Hossein
AH
Neuroscience Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran.
Sajadi
Mahnaz
M
Department of Biology, Tofigh Daru Co., Tehran, Iran.
Zohri
Mastaneh
M
Department of Biology, Tofigh Daru Co., Tehran, Iran.
Moosavi
Maryam
M
Department of Physiology and Neuroscience Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Zeinoddini
Mehdi
M
School of basic science, Tarbiat Modares University, Tehran, Iran.
D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D003384:Coxsackievirus Infections / Q000188:drug therapy / Q000821:virology*; D003546:Cysteine Endopeptidases / Q000502:physiology*; D029822:Enterovirus B, Human / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D010455:Peptides / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D013752:Tetracycline; D014764:Viral Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D014779:Virus Replication / Q000187:drug effects
D010455:Peptides; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus; D013752:Tetracycline