Journal article
A Quantitative and Kinetic Fusion Protein-Triggering Assay Can Discern Distinct Steps in the Nipah Virus Membrane Fusion Cascade
Journal of virology, Vol.84(16), pp.8033-8041
08/2010
Handle:
https://hdl.handle.net/2376/115241
PMCID: PMC2916531
PMID: 20519383
Abstract
The deadly paramyxovirus Nipah virus (NiV) contains a fusion glycoprotein (F) with canonical structural and functional features common to its class. Receptor binding to the NiV attachment glycoprotein (G) triggers F to undergo a two-phase conformational cascade: the first phase progresses from a metastable prefusion state to a prehairpin intermediate (PHI), while the second phase is marked by transition from the PHI to the six-helix-bundle hairpin. The PHI can be captured with peptides that mimic F's heptad repeat regions, and here we utilized a NiV heptad repeat peptide to quantify PHI formation and the half-lives (
t
1/2
) of the first and second fusion cascade phases. We found that ephrinB2 receptor binding to G triggered ∼2-fold more F than that triggered by ephrinB3, consistent with the increased rate and extent of fusion observed with ephrinB2- versus ephrinB3-expressing cells. In addition, for a series of hyper- and hypofusogenic F mutants, we quantified F-triggering capacities and measured the kinetics of their fusion cascade phases. Hyper- and hypofusogenicity can each be manifested through distinct stages of the fusion cascade, giving rise to vastly different half-lives for the first (
t
1/2
, 1.9 to 7.5 min) or second (
t
1/2
, 1.5 to 15.6 min) phase. While three mutants had a shorter first phase and a longer second phase than the wild-type protein, one mutant had the opposite phenotype. Thus, our results reveal multiple critical parameters that govern the paramyxovirus fusion cascade, and our assays should help efforts to elucidate other class I membrane fusion processes.
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Details
- Title
- A Quantitative and Kinetic Fusion Protein-Triggering Assay Can Discern Distinct Steps in the Nipah Virus Membrane Fusion Cascade
- Creators
- Hector C Aguilar - Department of Microbiology, Immunology and Molecular GeneticsVanessa Aspericueta - Department of Microbiology, Immunology and Molecular GeneticsLindsey R Robinson - Department of Microbiology, Immunology and Molecular GeneticsKaren E Aanensen - Department of Microbiology, Immunology and Molecular GeneticsBenhur Lee - Department of Microbiology, Immunology and Molecular Genetics
- Publication Details
- Journal of virology, Vol.84(16), pp.8033-8041
- Academic Unit
- Paul G. Allen School for Global Animal Health
- Publisher
- American Society for Microbiology (ASM)
- Number of pages
- 9
- Grant note
- Beckman Scholars Program UCLA AIDS Institute AI060694; AI069317; AI065359; AI07495; CA016042; AI028697 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA P30CA016042 / NATIONAL CANCER INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI) U54AI065359 / NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Allergy & Infectious Diseases (NIAID)
- Identifiers
- 99900547570101842
- Language
- English
- Resource Type
- Journal article