BACKGROUND INFORMATION: Protein-mediated merger of biological membranes, membrane fusion, is an important process. To investigate the role of fusogenic proteins in the initial size and dynamics of the fusion pore (a narrow aqueous pathway, which widens to finalize membrane fusion), two different fusion proteins expressed in the same cell line were investigated: the major glycoprotein of baculovirus Autographa californica (GP64) and the HA (haemagglutinin) of influenza X31. RESULTS: The host Sf9 cells expressing these viral proteins, irrespective of protein species, fused to human RBCs (red blood cells) upon acidification of the medium. A high-time-resolution electrophysiological study of fusion pore conductance revealed fundamental differences in (i) the initial pore conductance; pores created by HA were smaller than those created by GP64; (ii) the ability of pores to flicker; only HA-mediated pores flickered; and (iii) the time required for pore formation; HA-mediated pores took much longer to form after acidification. CONCLUSION: HA and GP64 have divergent electrophysiological phenotypes even when they fuse identical membranes, and fusion proteins play a crucial role in determining initial fusion pore characteristics. The structure of the initial fusion pore detected by electrical conductance measurements is sensitive to the nature of the fusion protein.
BACKGROUND INFORMATION: Protein-mediated merger of biological membranes, membrane fusion, is an important process. To investigate the role of fusogenic proteins in the initial size and dynamics of the fusion pore (a narrow aqueous pathway, which widens to finalize membrane fusion), two different fusion proteins expressed in the same cell line were investigated: the major glycoprotein of baculovirus Autographa californica (GP64) and the HA (haemagglutinin) of influenza X31. RESULTS: The host Sf9 cells expressing these viral proteins, irrespective of protein species, fused to human RBCs (red blood cells) upon acidification of the medium. A high-time-resolution electrophysiological study of fusion pore conductance revealed fundamental differences in (i) the initial pore conductance; pores created by HA were smaller than those created by GP64; (ii) the ability of pores to flicker; only HA-mediated pores flickered; and (iii) the time required for pore formation; HA-mediated pores took much longer to form after acidification. CONCLUSION: HA and GP64 have divergent electrophysiological phenotypes even when they fuse identical membranes, and fusion proteins play a crucial role in determining initial fusion pore characteristics. The structure of the initial fusion pore detected by electrical conductance measurements is sensitive to the nature of the fusion protein.
Authors: Don L Gibbons; Marie-Christine Vaney; Alain Roussel; Armelle Vigouroux; Brigid Reilly; Jean Lepault; Margaret Kielian; Félix A Rey Journal: Nature Date: 2004-01-22 Impact factor: 49.962
Authors: Loredana Vaccaro; Karen J Cross; Jens Kleinjung; Suzana K Straus; David J Thomas; Stephen A Wharton; John J Skehel; Franca Fraternali Journal: Biophys J Date: 2004-10-08 Impact factor: 4.033
Authors: Chang Sup Kim; Raquel F Epand; Eugenia Leikina; Richard M Epand; Leonid V Chernomordik Journal: J Biol Chem Date: 2011-02-03 Impact factor: 5.157