Closed and Semiclosed Interhelical Structures in Membrane vs Closed and Open Structures in Detergent for the Influenza Virus Hemagglutinin Fusion Peptide and Correlation of Hydrophobic Surface Area with Fusion Catalysis

Ghosh, Ujjayini; Xie, Long; Jia, Lihui; Liang, Shuang; Weliky, David P.

doi:10.1021/jacs.5b04578

Cited by 26 publications

(39 citation statements)

References 30 publications

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“…A ~65% α helical content is estimated from the experimental |θ 222 | ≈ 2 × 10 4 deg-cm 2 /dmole-residue and agrees semi-quantitatively with ~60 % α helical content calculated for HA2 based on the high-resolution structures of FP and SE fragments and the predicted α helical structure in the ~25-residue TM domain. 23,25,27 …”

Section: Resultsmentioning

confidence: 99%

“…27 Fusion is moderately higher for the 23-residue relative to the 20-residue peptide and for pH 5 relative to pH 7. Fusion for both peptides at both pH’s is difficult to understand based on the very different structures reported in detergent but is well-correlated to their structures in membrane.…”

Section: Introductionmentioning

confidence: 93%

“…Both peptides adopt ( N -helix)-(tight turn)-( C -helix) structure in membrane at both pH 5 and 7. 27–29 The helices can either pack tightly in the closed structure or a little less tightly in a semi-closed structure. For both peptides at both pH’s, there are significant populations of both structures with greater semi-closed population at pH 5.…”

Section: Introductionmentioning

confidence: 99%

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Full-length trimeric influenza virus hemagglutinin II membrane fusion protein and shorter constructs lacking the fusion peptide or transmembrane domain: Hyperthermostability of the full-length protein and the soluble ectodomain and fusion peptide make significant contributions to fusion of membrane vesicles

Ratnayake

Ekanayaka

Komanduru

et al. 2016

Protein Expression and Purification

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Influenza virus is a Class I enveloped virus which is initially endocytosed into a host respiratory epithelial cell. Subsequent reduction of the pH to the 5–6 range triggers a structural change of the viral hemagglutinin II (HA2) protein, fusion of the viral and endosomal membranes, and release of the viral nucleocapsid into the cytoplasm. HA2 contains fusion peptide (FP), soluble ectodomain (SE), transmembrane (TM), and intraviral domains with respective lengths of ~25, ~160, ~25, and ~10 residues. The present work provides a straightforward protocol for producing and purifying mg quantities of full-length HA2 from expression in bacteria. Biophysical and structural comparisons are made between full-length HA2 and shorter constructs including SHA2 ≡ SE, FHA2 ≡ FP + SE, and SHA2-TM ≡ SE + TM constructs. The constructs are helical in detergent at pH 7.4 and the dominant trimer species. The proteins are highly thermostable in decylmaltoside detergent with Tm > 90 °C for HA2 with stabilization provided by the SE, FP, and TM domains. The proteins are likely in a trimer-of-hairpins structure, the final protein state during fusion. All constructs induce fusion of negatively-charged vesicles at pH 5.0 with much less fusion at pH 7.4. Attractive protein/vesicle electrostatics play a role in fusion, as the proteins are positively-charged at pH 5.0 and negatively-charged at pH 7.4 and the pH-dependence of fusion is reversed for positively-charged vesicles. Comparison of fusion between constructs supports significant contributions to fusion from the SE and the FP with little effect from the TM.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Full-length trimeric influenza virus hemagglutinin II membrane fusion protein and shorter constructs lacking the fusion peptide or transmembrane domain: Hyperthermostability of the full-length protein and the soluble ectodomain and fusion peptide make significant contributions to fusion of membrane vesicles

Ratnayake

Ekanayaka

Komanduru

et al. 2016

Protein Expression and Purification

Self Cite

View full text Add to dashboard Cite

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“…Note, influenza hemagglutinin FPs showed large structural differences depending on the length of the amino acid sequences [24,25]. A recent study, using solid state NMR in lipid bilayer, has deduced close and semi-closed structural states of the hemagglutinin FP [72]. The determination of an amphipathic helical structure at the N-terminus of FP, in DPC micelle, may be considered as a novel feature of the SARS-CoV fusion protein.…”

Section: Discussionmentioning

confidence: 99%

NMR structure and localization of a large fragment of the SARS-CoV fusion protein: Implications in viral cell fusion

Mahajan

Chatterjee

Kannaian

et al. 2018

Biochimica et Biophysica Acta (BBA) - Biomembranes

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The lethal Coronaviruses (CoVs), Severe Acute Respiratory Syndrome-associated Coronavirus (SARS-CoV) and most recently Middle East Respiratory Syndrome Coronavirus, (MERS-CoV) are serious human health hazard. A successful viral infection requires fusion between virus and host cells carried out by the surface spike glycoprotein or S protein of CoV. Current models propose that the S2 subunit of S protein assembled into a hexameric helical bundle exposing hydrophobic fusogenic peptides or fusion peptides (FPs) for membrane insertion. The N-terminus of S2 subunit of SARS-CoV reported to be active in cell fusion whereby FPs have been identified. Atomic-resolution structure of FPs derived either in model membranes or in membrane mimic environment would glean insights toward viral cell fusion mechanism. Here, we have solved 3D structure, dynamics and micelle localization of a 64-residue long fusion peptide or LFP in DPC detergent micelles by NMR methods. Micelle bound structure of LFP is elucidated by the presence of discretely folded helical and intervening loops. The C-terminus region, residues F42-Y62, displays a long hydrophobic helix, whereas the N-terminus is defined by a short amphipathic helix, residues R4-Q12. The intervening residues of LFP assume stretches of loops and helical turns. The N-terminal helix is sustained by close aromatic and aliphatic sidechain packing interactions at the non-polar face. N{H}NOE studies indicated dynamical motion, at ps-ns timescale, of the helices of LFP in DPC micelles. PRE NMR showed that insertion of several regions of LFP into DPC micelle core. Together, the current study provides insights toward fusion mechanism of SARS-CoV.

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“…For example, certain single amino-acid substitutions in the fusion peptide abolish fusion, while not hampering HA expression or conformational changes (reviewed in [30]). On the membrane surface, the 23 amino-acid fusion peptide forms a helical hairpin structure [67,68] with an inverted wedge shape that induces negative curvature in the membrane [69]. This membrane deformation is thought to have a stabilizing effect on the hemifusion stalk with its strong negative curvature.…”

Section: Membrane Sculpting and Pore Formationmentioning

confidence: 99%

Mechanisms of influenza viral membrane fusion

Blijleven

Boonstra

Onck

et al. 2016

Seminars in Cell & Developmental Biology

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Influenza viral particles are enveloped by a lipid bilayer. A major step in infection is fusion of the viral and host cellular membranes, a process with large kinetic barriers. Influenza membrane fusion is catalyzed by hemagglutinin (HA), a class I viral fusion protein activated by low pH. The exact nature of the HA conformational changes that deliver the energy required for fusion remains poorly understood. This review summarizes our current knowledge of HA structure and dynamics, describes recent single-particle experiments and modeling studies, and discusses their role in understanding how multiple HAs mediate fusion. These approaches provide a mechanistic picture in which HAs independently and stochastically insert into the target membrane, forming a cluster of HAs that is collectively able to overcome the barrier to membrane fusion. The new experimental and modeling approaches described in this review hold promise for a more complete understanding of other viral fusion systems and the protein systems responsible for cellular fusion. Disciplines Medicine and Health Sciences | Social and Behavioral Sciences Publication DetailsBlijleven, J. S., Boonstra, S., Onck, P. R., van AbstractInfluenza viral particles are enveloped by a lipid bilayer. A major step in infection is fusion of the viral and host cellular membranes, a process with large kinetic barriers. Influenza membrane fusion is catalyzed by hemagglutinin (HA), a class I viral fusion protein activated by low pH. The exact nature of the HA conformational changes that deliver the energy required for fusion remains poorly understood. This review summarizes our current knowledge of HA structure and dynamics, describes recent single-particle experiments and modeling studies, and discusses their role in understanding how multiple HAs mediate fusion. These approaches provide a mechanistic picture in which HAs independently and stochastically insert into the target membrane, forming a cluster of HAs that is collectively able to overcome the barrier to membrane fusion. The new experimental and modeling approaches described in this review hold promise for a more complete understanding of other viral fusion systems and the protein systems responsible for cellular fusion.

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Closed and Semiclosed Interhelical Structures in Membrane vs Closed and Open Structures in Detergent for the Influenza Virus Hemagglutinin Fusion Peptide and Correlation of Hydrophobic Surface Area with Fusion Catalysis

Cited by 26 publications

References 30 publications

NMR structure and localization of a large fragment of the SARS-CoV fusion protein: Implications in viral cell fusion

Mechanisms of influenza viral membrane fusion

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