Peptides from conserved regions of paramyxovirus fusion (F) proteins are potent inhibitors of viral fusion.

Lambert, Dennis M.; Barney, Shawn; Lambert, A; Guthrie, Kelly; Medinas, R; Davis, Deodate; Bucy, Teresa B.; Erickson, Joel B.; Merutka, Gene; Petteway, Stephen R.

doi:10.1073/pnas.93.5.2186

Cited by 317 publications

(313 citation statements)

References 38 publications

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“…However, at a concentration of 10 g/ml or 2.8 M, the GP64 peptide inhibits slightly Ͻ50% of Sf9-RBC fusion and Sf9 cell-to-cell fusion. This concentration is several orders of magnitude greater than the 50% syncytial inhibition value of 100 pM reported for DP178 inhibition of HIV-mediated cell-to-cell fusion (Wild et al, 1994b) but may be closer to values reported for paramyxovirus fusion, which range from 0.015 to 2.1 M (Lambert et al, 1996;Yao and Compans, 1996).…”

Section: Discussionsupporting

confidence: 58%

“…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%

“…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: 91%

“…Examples include peptides derived from envelope proteins of HIV (Wild et al, , 1994b Jiang 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: Discussionsupporting

confidence: 58%

Section: The Heptad Repeat-dependent Stage Of Membrane Fusionmentioning

confidence: 99%

mentioning

confidence: 91%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A Discrete Stage of Baculovirus GP64-mediated Membrane Fusion

Kingsley

Behbahani

Rashtian

et al. 1999

MBoC

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“…To date, the majority of research into the fusion function of RSV F has been focused on regions within the F 1 subunit, in particular, the HRA and HRB coiled coil (23)(24)(25). While these studies have provided deeper insight into formation and antiviral targeting of the terminal 6HB fusion core (26), the roles of residues and domains within the F 2 subunit remain relatively unexplored.…”

Section: Discussionmentioning

confidence: 99%

The Heptad Repeat C Domain of the Respiratory Syncytial Virus Fusion Protein Plays a Key Role in Membrane Fusion

Bermingham

Chappell

Watterson

et al. 2018

J Virol

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Respiratory syncytial virus (RSV) mediates host cell entry through the fusion (F) protein, which undergoes a conformational change to facilitate the merger of viral and host lipid membrane envelopes. The RSV F protein comprises a trimer of disulfide-bonded F 1 and F 2 subunits that is present on the virion surface in a metastable prefusion state. This prefusion form is readily triggered to undergo refolding to bring two heptad repeats (heptad repeat A [HRA] and HRB) into close proximity to form a six-helix bundle that stabilizes the postfusion form and provides the free energy required for membrane fusion. This process can be triggered independently of other proteins. Here, we have performed a comprehensive analysis of a third heptad repeat region, HRC (amino acids 75 to 97), an amphipathic ␣-helix that lies at the interface of the prefusion F trimer and is a major structural feature of the F 2 subunit. We performed alanine scanning mutagenesis from Lys-75 to Met-97 and assessed all mutations in transient cell culture for expression, proteolytic processing, cell surface localization, protein conformation, and membrane fusion. Functional characterization revealed a striking distribution of activity in which fusion-increasing mutations localized to one side of the helical face, while fusion-decreasing mutations clustered on the opposing face. Here, we propose a model in which HRC plays a stabilizing role within the globular head for the prefusion F trimer and is potentially involved in the early events of triggering, prompting fusion peptide release and transition into the postfusion state.IMPORTANCE RSV is recognized as the most important viral pathogen among pediatric populations worldwide, yet no vaccine or widely available therapeutic treatment is available. The F protein is critical for the viral replication process and is the major target for neutralizing antibodies. Recent years have seen the development of prefusion stabilized F protein-based approaches to vaccine design. A detailed understanding of the specific domains and residues that contribute to protein stability and fusion function is fundamental to such efforts. Here, we present a comprehensive mutagenesis-based study of a region of the RSV F 2 subunit (amino acids 75 to 97), referred to as HRC, and propose a role for this helical region in maintaining the delicate stability of the prefusion form.KEYWORDS respiratory syncytial virus, fusion, membrane fusion, mutagenesis, heptad repeat R espiratory syncytial virus (RSV) is widely considered the most significant viral pediatric pathogen worldwide. Belonging to the Pneumovirinae family, RSV causes substantial morbidity and mortality worldwide, with an estimated 33.8 million acute lower respiratory tract infections per year in children under 5 years of age, and has been linked to almost 200,000 deaths annually, 99% of which are in developing countries (1, 2). RSV is also recognized as an important pathogen of high-risk adult and elderly populations (3). Despite this, no targeted drug or vaccin...

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Antiviral Agents

Hosmane

2003

Kirk-Othmer Encyclopedia of Chemical Technology

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Since the discovery of the first virus >100 years ago, thousands of different viruses have been identified and characterized, and a number of plant, animal, and human ailments have been traced to viral origin. The rapid advances made in tools and techniques of molecular biology in the last 40 years, coupled with cooperative efforts in genetics and biochemical fronts, have afforded intricate details of the structure, function, replication, and genomic makeup of a host of viruses. As there are too many viruses to give even short accounts, this article focuses on only those that have some relevance to human diseases. But, since the list of all human viruses would still be too long to give detailed descriptions for each, the focus is further narrowed down to four major viruses that are currently perceived to threaten global health. These four viruses include HIV (human immunodeficiency virus, which causes AIDS), HBV and HCV (hepatitis B and C viruses that cause liver damage), and the more recent WNV (West Nile virus that causes brain inflammation). Nevertheless, an attempt has been made to classify all the known major viruses based on their shape, size, symmetry, hosts, and chemical composition that includes nucleic acid, protein, and presence or absence of lipid envelope. As they are a form of life that cannot replicate outside a host cell, viruses have evolved to develop complex and diverse interactions with higher organisms. Therefore, it was long believed that the development of antiviral agents that would specifically disrupt the viral replication process without affecting the normal metabolic events of the host would be difficult. Thanks to excellent advances in virology, several targets that are unique to the viruses have now been identified and successfully explored. This article briefly describes the general processes of viral infection to point out potential targets for selective antiviral agents. These targets are then classified into two major virus‐specific processes, including ( a ) early events of viral adsorption, penetration and uncoating, and ( b ) later synthetic events that concern intracellular replication of the virus. While there are only limited choices of candidates dealing with the early events, a much larger pool of candidates exists for targeting the later events in a virus life cycle, and they are known to be virus‐specific. The viruses synthesize and utilize specific enzymes and proteins, and more importantly, the replication of viral genomes is also virus‐specific. Nucleoside analogues which target these later events of viral life cycles have played major roles as antiviral agents against almost all the major viruses, and therefore, a special emphasis has been placed on this class of compounds in this review article. Prophylactic measures such as vaccine immunization have also been discussed under each of the four major viruses mentioned.

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Peptides from conserved regions of paramyxovirus fusion (F) proteins are potent inhibitors of viral fusion.

Cited by 317 publications

References 38 publications

A Discrete Stage of Baculovirus GP64-mediated Membrane Fusion

A Discrete Stage of Baculovirus GP64-mediated Membrane Fusion

The Heptad Repeat C Domain of the Respiratory Syncytial Virus Fusion Protein Plays a Key Role in Membrane Fusion

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