Implication of the proprotein convertases furin, PC5 and PC7 in the cleavage of surface glycoproteins of Hong Kong, Ebola and respiratory syncytial viruses: a comparative analysis with fluorogenic peptides.
Fluorogenic peptides encompassing the processing sites of envelope glycoproteins of the infectious influenza A Hong Kong virus (HKV), Ebola virus (EBOV) and respiratory syncytial virus (RSV) were tested for cleavage by soluble recombinants of the proprotein convertases furin, PC5 and PC7. Kinetic studies with these intramolecularly quenched fluorogenic peptides revealed selective cleavages at the physiological dibasic sites. The HKV peptide is cleaved by both furin and PC5 with similar efficacy; in comparison, PC7 cleaves this substrate poorly. In contrast with the basic tetrapeptide insertion within the haemagglutinin sequence of HKV, two other dipeptide insertions revealed a poorer cleavage with a similar rank order of potency. These results demonstrate that the N-terminal RERR insertion to the wild-type avian RKKR downward arrow sequence is functionally significant, and suggest that the approx. 5-fold increase in cleavage efficacy contributes to the high infectivity of the H5N1 virus subtype. With regard to RSV peptide processing, PC7 is twice as effective as PC5 and furin. The EBOV peptide was processed with similar efficiency by the three enzymes. Our observations that all of these cleavages can be effectively inhibited by a plant andrographolide derivative at 250 microM or less might aid in the design of potent convertase inhibitors as alternative antiviral therapies.
Basak
A
A
Laboratory of Molecular Medicine and Disease of Ageing Centre, Loeb Health Research Institute, Ottawa Civic Hospital, 725 Parkdale Avenue, Ottawa, Ontario, Canada K1Y 4K9. abasak@lri.ca
Zhong
M
M
Munzer
J S
JS
Chrétien
M
M
Seidah
N G
NG
D000595:Amino Acid Sequence; D029043:Ebolavirus / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D045683:Furin; D006868:Hydrolysis; D009980:Influenza A virus / Q000378:metabolism*; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012136:Respiratory Syncytial Viruses / Q000378:metabolism*; D013381:Subtilisins / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism
D005456:Fluorescent Dyes; D008562:Membrane Glycoproteins; D010455:Peptides; D011994:Recombinant Proteins; D014764:Viral Proteins; C099101:PCSK7 protein, human; D013381:Subtilisins; D045683:Furin