H+-induced membrane insertion of influenza virus hemagglutinin involves the HA2 amino-terminal fusion peptide but not the coiled coil region.
Fusion of influenza virus with target membranes is induced by acid and involves complex changes in the viral envelope protein hemagglutinin (HA). In a first, kinetically distinct step, the HA polypeptide chain 2 (HA2) is inserted into the target membrane bilayer. Using hydrophobic photolabeling with the phospholipid analogue 1-O-hexadecanoyl-2-O-[9-[[[2-[125I]iodo-4(trifluoromethyl-3H-diazirin -3-yl)benzyl]oxy]carbonyl]nonanoyl]-sn-glycero-3-phosphocholine, we identified the segment within HA2 that interacts with the membrane. The sole part of the HA2 ectodomain that was labeled with the membrane-restricted reagent is the NH2-terminal fusion peptide (residues 1-22). No labeling occurred within the long coiled coil region generated during the acid-induced conformational transition (Bullough, P. A., Hughson, F. M., Skehel, J. J., and Wiley, D. C. (1994) Nature 371, 37-43). These data strongly suggest that the coiled coil region of HA2 does not insert into the lipid bilayer. This conclusion is at variance with the recent suggestion (Yu, Y. G., King, D. S., and Shin, Y.-K.(1994) Science 266, 274-276) that the coiled coil of HA may splay apart and insert into the target membrane, providing a mechanism by which the viral and the target membrane may come in close apposition.
Durrer
P
P
Laboratory of Biochemistry, Eidgenössische Technische Hochschule Zürich (ETHZ), ETH-Zentrum, CH-8092 Zürich, Switzerland.
Galli
C
C
Hoenke
S
S
Corti
C
C
Glück
R
R
Vorherr
T
T
Brunner
J
J
D000818:Animals; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000737:chemistry* / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008081:Liposomes; D008561:Membrane Fusion / Q000502:physiology; D008954:Models, Biological; D015394:Molecular Structure; D009975:Orthomyxoviridae / Q000737:chemistry / Q000502:physiology*; D011487:Protein Conformation; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000502:physiology
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D008081:Liposomes; D014760:Viral Fusion Proteins