Isolation, identification and function of a novel anti-HSV-1 protein from Grifola frondosa.
A novel antiviral protein was purified from an extract of Grifola frondosa fruiting bodies using a procedure that included 40% ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography, and designated GFAHP. This protein inhibited herpes simplex virus type 1 (HSV-1) replication in vitro with an IC(50) value of 4.1 microg/ml and a therapeutic index >29.3. Higher concentrations of GFAHP (125 and 500 microg/ml) also significantly reduced the severity of HSV-1 induced blepharitis, neovascularization, and stromal keratitis in a murine model. Topical administration of GFAHP to the mouse cornea resulted in a significant decrease in virus production (mean virus yields: 3.4log10PFU in the treated group and 4.19log10PFU in the control group). We proved that GFAHP directly inactivates HSV-1 while simultaneously inhibiting HSV-1 penetration into Vero cells. Gel electrophoresis showed that GFAHP had a molecular weight of 29.5 kDa. GFAHP was tryptic digested and analyzed from the PMF of matrix assisted desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) and nanoelectrospray ionization tandem mass spectrometry. The N-terminal sequence of GFAHP consisted of an 11 amino acid peptide, NH(2)-REQDNAPCGLN-COOH that did not match any known amino acid sequences, indicating that GFAHP is likely to be a novel antivirus protein. To our knowledge, this is the first report that characterizes an anti-HSV protein from G. frondosa.
Gu
Chang-Qing
CQ
Institute of Environment and Health, 1 Dali Road, Tianjin 300050, PR China.
Li
Jun-Wen
JW
Chao
Fuhuan
F
Jin
Min
M
Wang
Xin-Wei
XW
Shen
Zhi-Qiang
ZQ
D000818:Animals; D000998:Antiviral Agents / Q000302:isolation & purification* / Q000494:pharmacology / Q000627:therapeutic use; D002460:Cell Line; D003315:Cornea / Q000821:virology; D005260:Female; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology / Q000627:therapeutic use*; D048691:Grifola / Q000737:chemistry*; D006561:Herpes Simplex / Q000188:drug therapy* / Q000821:virology; D018259:Herpesvirus 1, Human / Q000187:drug effects* / Q000502:physiology; D016849:Keratitis, Herpetic / Q000188:drug therapy* / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D005656:Fungal Proteins