Inhibition of parainfluenza virus type 3 and Newcastle disease virus hemagglutinin-neuraminidase receptor binding: effect of receptor avidity and steric hindrance at the inhibitor binding sites.
Zanamivir (4-guanidino-Neu5Ac2en [4-GU-DANA]) inhibits not only the neuraminidase activity but also the receptor interaction of the human parainfluenza virus type 3 (HPIV3) hemagglutinin-neuraminidase (HN), blocking receptor binding and subsequent fusion promotion. All activities of the HPIV3 variant ZM1 HN (T193I/I567V) are less sensitive to 4-GU-DANA's effects. The T193I mutation in HN confers both increased receptor binding and increased neuraminidase activity, as well as reduced sensitivities of both activities to 4-GU-DANA inhibition, consistent with a single site on the HN molecule carrying out both catalysis and binding. We now provide evidence that the HPIV3 variant's resistance to receptor-binding inhibition by 4-GU-DANA is related to a reduced affinity of the HN receptor-binding site for this compound as well as to an increase in the avidity of HN for the receptor. Newcastle disease virus (NDV) HN and HPIV3 HN respond differently to inhibition in ways that suggest a fundamental distinction between them. NDV HN-receptor binding is less sensitive than HPIV3 HN-receptor binding to 4-GU-DANA, while its neuraminidase activity is highly sensitive. Both HPIV3 and NDV HNs are sensitive to receptor-binding inhibition by the smaller molecule DANA. However, for NDV HN, some receptor binding cannot be inhibited. These data are consistent with the presence in NDV HN of a second receptor-binding site that is devoid of enzyme activity and has a negligible, if any, affinity for 4-GU-DANA. Avidity for the receptor contributes to resistance by allowing the receptor to compete effectively with inhibitors for interaction with HN, while the further determinant of resistance is the reduced binding of the inhibitor molecule to the binding pocket on HN. Based upon our data and recent three-dimensional structural information on the HPIV3 and NDV HNs, we propose mechanisms for the observed sensitivity and resistance of HN to receptor-binding inhibition and discuss the implications of these mechanisms for the distribution of HN functions.
Porotto
Matteo
M
Department of Pediatrics, Mount Sinai School of Medicine, 1 Gustave L. Levy Pl., New York, NY 10029, USA.
Murrell
Matthew
M
Greengard
Olga
O
Lawrence
Michael C
MC
McKimm-Breschkin
Jennifer L
JL
Moscona
Anne
A
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D001665:Binding Sites; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006146:Guanidines; D015707:HN Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D009154:Mutation; D009439:Neuraminidase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009522:Newcastle disease virus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D010224:Parainfluenza Virus 3, Human / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D011714:Pyrans; D011991:Receptors, Virus / Q000737:chemistry / Q000378:metabolism*; D012794:Sialic Acids / Q000378:metabolism / Q000494:pharmacology*; D053243:Zanamivir
D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D006146:Guanidines; D015707:HN Protein; C039111:HN protein receptor; D011714:Pyrans; D011991:Receptors, Virus; D012794:Sialic Acids; D009439:Neuraminidase; D053243:Zanamivir