scholarly journals Development of a high-throughput assay to detect antibody inhibition of low pH induced conformational changes of influenza virus hemagglutinin

PLoS ONE ◽  
2018 ◽  
Vol 13 (6) ◽  
pp. e0199683 ◽  
Author(s):  
Jessica F. Trost ◽  
Elizabeth H. LeMasters ◽  
Feng Liu ◽  
Paul Carney ◽  
Xiuhua Lu ◽  
...  
Keyword(s):  
Influenza Virus ◽  
High Throughput ◽  
Conformational Changes ◽  
Low Ph ◽  
Influenza Virus Hemagglutinin ◽  
High Throughput Assay ◽  
Antibody Inhibition

scholarly journals Anti-peptide antibodies detect steps in a protein conformational change: low-pH activation of the influenza virus hemagglutinin.

1987 ◽  
Vol 105 (6) ◽  
pp. 2887-2896 ◽  
Author(s):  
J M White ◽  
I A Wilson
Keyword(s):  
Influenza Virus ◽  
Membrane Fusion ◽  
Conformational Change ◽  
Conformational Changes ◽  
Three Dimensional ◽  
Low Ph ◽  
Protein Conformational Changes ◽  
Influenza Virus Hemagglutinin ◽  
Protein Conformational Change ◽  
Peptide Antibodies

At low pH, the hemagglutinin (HA) of influenza virus undergoes an irreversible conformational change that potentiates its essential membrane fusion function. We have probed the details of this conformational change using a panel of 14 anti-HA-peptide antibodies. Whereas some antibodies reacted equally well with both the neutral and low-pH HA conformations, others reacted to a significantly greater extent with the low-pH form. The locations of the peptides recognized by the latter antibodies in the three-dimensional HA structure indicated regions of the protein that change in response to low pH. Moreover, kinetic experiments suggested steps in the conformational change. In addition to their relevance to membrane fusion, our results show that anti-peptide antibodies can be used to study some types of biologically important protein conformational changes.

scholarly journals New Insights into the Spring-Loaded Conformational Change of Influenza Virus Hemagglutinin

2002 ◽  
Vol 76 (9) ◽  
pp. 4456-4466 ◽  
Author(s):  
Jennifer A. Gruenke ◽  
R. Todd Armstrong ◽  
William W. Newcomb ◽  
Jay C. Brown ◽  
Judith M. White
Keyword(s):  
Influenza Virus ◽  
Conformational Change ◽  
Conformational Changes ◽  
Limited Proteolysis ◽  
Coiled Coil ◽  
Fusion Peptide ◽  
Wild Type ◽  
Influenza Virus Hemagglutinin ◽  
Target Membrane ◽  
Mutant Forms

ABSTRACT Influenza virus hemagglutinin undergoes a conformational change in which a loop-to-helix “spring-loaded” conformational change forms a coiled coil that positions the fusion peptide for interaction with the target bilayer. Previous work has shown that two proline mutations designed to disrupt this change disrupt fusion but did not determine the basis for the fusion defect. In this work, we made six additional mutants with single proline substitutions in the region that undergoes the spring-loaded conformational change and two additional mutants with double proline substitutions in this region. All double mutants were fusion inactive. We analyzed one double mutant, F63P/F70P, as an example. We observed that F63P/F70P undergoes key low-pH-induced conformational changes and binds tightly to target membranes. However, limited proteolysis and electron microscopy observations showed that the mutant forms a coiled coil that is only ∼50% the length of the wild type, suggesting that it is splayed in its N-terminal half. This work further supports the hypothesis that the spring-loaded conformational change is necessary for fusion. Our data also indicate that the spring-loaded conformational change has another role beyond presenting the fusion peptide to the target membrane.

Deacylation of influenza virus hemagglutinin does not affect the kinetics of low pH induced membrane fusion

1996 ◽  
Vol 431 (S6) ◽  
pp. R257-R258 ◽  
Author(s):  
Britta Schroth ◽  
Hans C. Philipp ◽  
Michael Veit ◽  
Michael F. G. Schmidt ◽  
Andreas Herrmann
Keyword(s):  
Influenza Virus ◽  
Membrane Fusion ◽  
Low Ph ◽  
Induced Membrane ◽  
Influenza Virus Hemagglutinin ◽  
Kinetics Of

scholarly journals Conformational Intermediates and Fusion Activity of Influenza Virus Hemagglutinin

1999 ◽  
Vol 73 (6) ◽  
pp. 4567-4574 ◽  
Author(s):  
Thomas Korte ◽  
Kai Ludwig ◽  
Frank P. Booy ◽  
Robert Blumenthal ◽  
Andreas Herrmann
Keyword(s):  
Influenza Virus ◽  
High Resolution ◽  
Low Ph ◽  
Fusion Activity ◽  
Native Structure ◽  
Ph Values ◽  
Influenza Virus Hemagglutinin ◽  
High Resolution Structure ◽  
H3 Subtype ◽  
Resolution Structure

ABSTRACT Three strains of influenza virus (H1, H2, and H3) exhibited similar characteristics in the ability of their hemagglutinin (HA) to induce membrane fusion, but the HAs differed in their susceptibility to inactivation. The extent of inactivation depended on the pH of preincubation and was lowest for A/Japan (H2 subtype), in agreement with previous studies (A. Puri, F. Booy, R. W. Doms, J. M. White, and R. Blumenthal, J. Virol. 64:3824–3832, 1990). While significant inactivation of X31 (H3 subtype) was observed at 37°C at pH values corresponding to the maximum of fusion (about pH 5.0), no inactivation was seen at preincubation pH values 0.2 to 0.4 pH units higher. Surprisingly, low-pH preincubation under those conditions enhanced the fusion rates and extents of A/Japan as well as those of X31. For A/PR 8/34 (H1 subtype), neither a shift of the pH (to >5.0) nor a decrease of the temperature to 20°C was sufficient to prevent inactivation. We provide evidence that the activated HA is a conformational intermediate distinct from the native structure and from the final structure associated with the conformational change of HA, which is implicated by the high-resolution structure of the soluble trimeric fragment TBHA2 (P. A. Bullough, F. M. Hughson, J. J. Skehel, and D. C. Wiley, Nature 371:37–43, 1994).

Spin-labeling of influenza virus hemagglutinin permits analysis of the conformational change at low pH and its inhibition by antibody

1986 ◽  
Vol 4 (3) ◽  
pp. 251-261 ◽  
Author(s):  
Sayaka Yoden ◽  
Hiroshi Kida ◽  
Mikinori Kuwabara ◽  
Ryo Yanagawa ◽  
Robert G. Webster
Keyword(s):  
Influenza Virus ◽  
Conformational Change ◽  
Spin Labeling ◽  
Low Ph ◽  
Influenza Virus Hemagglutinin

scholarly journals Class II fusion protein of alphaviruses drives membrane fusion through the same pathway as class I proteins

2005 ◽  
Vol 169 (1) ◽  
pp. 167-177 ◽  
Author(s):  
Elena Zaitseva ◽  
Aditya Mittal ◽  
Diane E. Griffin ◽  
Leonid V. Chernomordik
Keyword(s):  
Influenza Virus ◽  
Fusion Proteins ◽  
Low Ph ◽  
Viral Fusion ◽  
Fusion Reactions ◽  
Influenza Virus Hemagglutinin ◽  
Viral Fusion Proteins ◽  
The Difference ◽  
Fusion Pores ◽  
Specific Inhibitors

Viral fusion proteins of classes I and II differ radically in their initial structures but refold toward similar conformations upon activation. Do fusion pathways mediated by alphavirus E1 and influenza virus hemagglutinin (HA) that exemplify classes II and I differ to reflect the difference in their initial conformations, or concur to reflect the similarity in the final conformations? Here, we dissected the pathway of low pH–triggered E1-mediated cell–cell fusion by reducing the numbers of activated E1 proteins and by blocking different fusion stages with specific inhibitors. The discovered progression from transient hemifusion to small, and then expanding, fusion pores upon an increase in the number of activated fusion proteins parallels that established for HA-mediated fusion. We conclude that proteins as different as E1 and HA drive fusion through strikingly similar membrane intermediates, with the most energy-intensive stages following rather than preceding hemifusion. We propose that fusion reactions catalyzed by all proteins of both classes follow a similar pathway.

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