Please use this identifier to cite or link to this item: http://hdl.handle.net/10316/8436
Title: Kinetic modeling of Sendai virus fusion with PC-12 cells. Effect of pH and temperature on fusion and viral inactivation
Authors: Lima, Maria da Conceição Pedroso de 
Ramalho-Santos, João 
Martins, Maria de Fátima 
Carvalho, Arsélio Pato de 
Bairos, Vasco 
Nir, Shlomo 
Issue Date: 1992
Citation: European Journal of Biochemistry. 205:1 (1992) 181-186
Abstract: We have studied the fusion activity of Sendai virus, a lipid-enveloped paramyxovirus, towards a line of adherent cells designated PC-12. Fusion was monitored by the dequenching of octadecylrhodamine, a fluorescent non-exchangeable probe. The results were analysed with a mass action kinetic model which could explain and predict the kinetics of virus2013cell fusion. When the temperature was lowered from 37°C to 25°C, a sharp inhibition of the fusion process was observed, probably reflecting a constraint in the movement of viral glycoproteins at low temperatures. The rate constants of adhesion and fusion were reduced 3.5-fold and 7-fold, respectively, as the temperature was lowered from 37°C to 25°C. The fusion process seemed essentially pH-independent, unlike the case of liposomes and erythrocyte ghosts. Preincubation of the virus in the absence of target cell membranes at neutral and alkaline pH (37°C, 30 min) did not affect the fusion process. However, a similar preincubation of the virus at pH = 5.0 resulted in marked, though slow, inhibition in fusion with the fusion rate constant being reduced 8-fold. Viral preincubation for 5 min in the same acidic conditions yielded a mild inhibition of fusogenic activity, while preincubation in the cold (4°C, 30 min) did not alter viral fusion activity. These acid-induced inhibitory effects could not be fully reversed by further viral preincubation at pH = 7.4 (37°C, 30 min). Changes in internal pH as well as endocytic activity of PC-12 cells had small effect on the fusion process, thus indicating that Sendai virus fuses primarily with the plasma membranes.
URI: http://hdl.handle.net/10316/8436
DOI: 10.1111/j.1432-1033.1992.tb16766.x
Rights: openAccess
Appears in Collections:FMUC Medicina - Artigos em Revistas Internacionais

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