Analysis of a Peptide Inhibitor of Paramyxovirus (NDV) Fusion Using Biological Assays, NMR, and Molecular Modeling

Young, John K.; Hicks, Rickey P.; Wright, George E.; Morrison, Trudy G.

doi:10.1006/viro.1997.8834

Cited by 81 publications

(79 citation statements)

References 41 publications

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“…Hydrophobic 4-3 heptad repeat motifs are clearly important for pH-independent viral protein-mediated fusion (Chen et al, 1995;Reitter et al, 1995;Lambert et al, 1996;Young et al, 1997). For HIV gp41 peptide inhibition, it has been predicted that peptides would exert inhibitory effects during the conformational change to the fusion-active conformation (Wiessenhorn et al, 1997).…”

Section: The Heptad Repeat-dependent Stage Of Membrane Fusionmentioning

confidence: 99%

“…The frequent observation of hydrophobic 4-3 heptad repeats (leucine zipper-like) motifs within viral fusion proteins, coupled with the evidence for functional roles for both pH-independent and now pH-dependent viral fusion processes, suggests that the hydrophobic 4-3 heptad motif performs a universal function critical to membrane fusion in general. It should be noted, however, that all hydrophobic 4-3 heptad repeat regions of pH-independent fusion proteins may not function at the same stage of membrane fusion, because Young et al (1997) Exactly what specific functional role (or roles) the hydrophobic 4-3 heptad repeat plays during GP64-mediated membrane fusion also remains hypothetical. Indeed, it is not clear how many conformational intermediates ensue after the pH-driven triggering of GP64, nor which of the putative intermediate(s) is actually fusogenic.…”

Section: The Heptad Repeat-dependent Stage Of Membrane Fusionmentioning

confidence: 99%

“…Because two discrete HIV GP41 hydrophobic 4-3 heptad repeats peptides (DP107 and DP178) interact in solution, these two regions may interact to form a "molecular clamp" that stabilizes the inactive conformation of the GP120-GP41 complex (Chen et al, 1995). Experiments with paramyxovirus heptad repeat peptides also appear to support the molecular clamp hypothesis because (a) these peptides demonstrate strain-specific inhibition of F protein function and (b) the two 4-3 heptad repeat regions interact in solution (Lambert et al, 1996;Young et al, 1997Young et al, , 1999. It has also been suggested that the hydrophobic 4-3 heptad repeat sequences form a hydrophobic face that directly interacts with or disturbs the host membrane concomitant with disruption of the target membrane (Rabenstein and Shin, 1995;Reitter et al 1995).…”

mentioning

confidence: 90%

“…Examples include peptides derived from envelope proteins of HIV (Wild et al, , 1994bJiang et al, 1993a,b;Judice et al, 1997) and paramyxoviruses (Rapaport et al, 1995;Lambert et al, 1996;Yao and Compans, 1996;Ghosh et al, 1997;Wild and Buckland, 1997;Young et al, 1997;Ben-Efraim et al, 1999). These peptides are presumed to mimic functional domains of the paramyxovirus and HIV fusion proteins and therefore disrupt the activity of these proteins (Young et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

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A Discrete Stage of Baculovirus GP64-mediated Membrane Fusion

Kingsley

Behbahani

Rashtian

et al. 1999

MBoC

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Viral fusion protein trimers can play a critical role in limiting lipids in membrane fusion. Because the trimeric oligomer of many viral fusion proteins is often stabilized by hydrophobic 4-3 heptad repeats, higher-order oligomers might be stabilized by similar sequences. There is a hydrophobic 4-3 heptad repeat contiguous to a putative oligomerization domain of Autographa californica multicapsid nucleopolyhedrovirus envelope glycoprotein GP64. We performed mutagenesis and peptide inhibition studies to determine if this sequence might play a role in catalysis of membrane fusion. First, leucine-to-alanine mutants within and flanking the amino terminus of the hydrophobic 4-3 heptad repeat motif that oligomerize into trimers and traffic to insect Sf9 cell surfaces were identified. These mutants retained their wild-type conformation at neutral pH and changed conformation in acidic conditions, as judged by the reactivity of a conformationally sensitive mAb. These mutants, however, were defective for membrane fusion. Second, a peptide encoding the portion flanking the GP64 hydrophobic 4-3 heptad repeat was synthesized. Adding peptide led to inhibition of membrane fusion, which occurred only when the peptide was present during low pH application. The presence of peptide during low pH application did not prevent low pH-induced conformational changes, as determined by the loss of a conformationally sensitive epitope. In control experiments, a peptide of identical composition but different sequence, or a peptide encoding a portion of the Ebola GP heptad motif, had no effect on GP64-mediated fusion. Furthermore, when the hemagglutinin (X31 strain) fusion protein of influenza was functionally expressed in Sf9 cells, no effect on hemagglutinin-mediated fusion was observed, suggesting that the peptide does not exert nonspecific effects on other fusion proteins or cell membranes. Collectively, these studies suggest that the specific peptide sequences of GP64 that are adjacent to and include portions of the hydrophobic 4-3 heptad repeat play a dynamic role in membrane fusion at a stage that is downstream of the initiation of protein conformational changes but upstream of lipid mixing. INTRODUCTIONThe mechanism of membrane fusion is currently a field of intense investigation, both for intracellular trafficking and enveloped viral infection. For viral fusion protein function, we have previously suggested that fusion protein oligomers form transient and cooperative higher-order structures or rings . These transient ring structures are proposed to facilitate conversion of kinetic energy, released during protein conformational change, into work on the target and host membranes required for membrane fusion. Indeed, biochemical evidence for baculovirus GP64 aggregates during membrane fusion has recently been reported (Markovic et al. 1998). Whether and how viral fusion proteins might function in a coordinated and cooperative manner is a major question in the field of membrane fusion. Because hydrophobic 4-3 heptad repeats are observed...

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Section: The Heptad Repeat-dependent Stage Of Membrane Fusionmentioning

confidence: 99%

Section: The Heptad Repeat-dependent Stage Of Membrane Fusionmentioning

confidence: 99%

mentioning

confidence: 90%

Section: Introductionmentioning

confidence: 99%

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A Discrete Stage of Baculovirus GP64-mediated Membrane Fusion

Kingsley

Behbahani

Rashtian

et al. 1999

MBoC

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“…Synthetic peptides corresponding to both HR-1 and -2 of the HIV gp41 protein have been shown to inhibit viral fusion (31,40,41). Due to the relatively high potency of HR-2 peptides, this approach has been extended to peptides derived from the fusion proteins of several viruses including RSV (17), hPIV3 (17,43), avian pneumovirus (37), Newcastle disease virus (46), measles virus (17), and SV5 (25). It also has been noted that HR peptides are virus specific; no peptide has been shown to inhibit multiple viruses.…”

mentioning

confidence: 99%

Examination of a Fusogenic Hexameric Core from Human Metapneumovirus and Identification of a Potent Synthetic Peptide Inhibitor from the Heptad Repeat 1 Region

Miller

Crowe

Williams³

et al. 2007

J Virol

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Paramyxoviruses are a leading cause of childhood illness worldwide. A recently discovered paramyxovirus, human metapneumovirus (hMPV), has been studied by our group in order to determine the structural relevance of its fusion (F) protein to other well-characterized viruses utilizing type I integral membrane proteins as fusion aids. Sequence analysis and homology models suggested the presence of requisite heptad repeat (HR) regions. Synthetic peptides from HR regions 1 and 2 (HR-1 and -2, respectively) were induced to form a thermostable (melting temperature, ϳ90°C) helical structure consistent in mass with a hexameric coiled coil. Inhibitory studies of hMPV HR-1 and -2 indicated that the synthetic HR-1 peptide was a significant fusion inhibitor with a 50% inhibitory concentration and a 50% effective concentration of ϳ50 nM. Many viral fusion proteins are type I integral membrane proteins utilizing the formation of a hexameric coiled coil of HR peptides as a major driving force for fusion. Our studies provide evidence that hMPV also uses a coiled-coil structure as a major player in the fusion process. Additionally, viral HR-1 peptide sequences may need further investigation as potent fusion inhibitors.

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