VP4 Protein from Human Rhinovirus 14 Is Released by Pressure and Locked in the Capsid by the Antiviral Compound WIN

Rafael B. Gonçalves; Ygara S. Mendes; Marcia R. Soares; Umesh Katpally; Thomas J. Smith; Jerson L. Silva; Andréa C. Oliveira

doi:10.1016/j.jmb.2006.11.033

VP4 Protein from Human Rhinovirus 14 Is Released by Pressure and Locked in the Capsid by the Antiviral Compound WIN

Rafael B. Gonçalves, Ygara S. Mendes, Marcia R. Soares, Umesh Katpally, Thomas J. Smith, Jerson L. Silva, Andréa C. Oliveira

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

Rhinoviruses are the major causative agents of the common cold in humans. Here, we studied the stability of human rhinovirus type 14 (HRV14) under conditions of high hydrostatic pressure, low temperature, and urea in the absence and presence of an antiviral drug. Capsid dissociation and changes in the protein conformation were monitored by fluorescence spectroscopy, light scattering, circular dichroism, gel filtration chromatography, mass spectrometry and infectivity assays. The data show that high pressure induces the dissociation of HRV14 and that this process is inhibited by WIN 52084. MALDI-TOF mass spectrometry experiments demonstrate that VP4, the most internal viral protein, is released from the capsid by pressure treatment. This release of VP4 is concomitant with loss of infectivity. Our studies also show that at least one antiviral effect of the WIN drugs involves the locking of VP4 inside the capsid by blocking the dynamics associated with cell attachment.

Original language	English (US)
Pages (from-to)	295-306
Number of pages	12
Journal	Journal of Molecular Biology
Volume	366
Issue number	1
DOIs	https://doi.org/10.1016/j.jmb.2006.11.033
State	Published - Feb 9 2007
Externally published	Yes

Keywords

WIN drugs
fluorescence
human rhinovirus
hydrostatic pressure
mass spectrometry

ASJC Scopus subject areas

Biophysics
Structural Biology
Molecular Biology

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10.1016/j.jmb.2006.11.033

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title = "VP4 Protein from Human Rhinovirus 14 Is Released by Pressure and Locked in the Capsid by the Antiviral Compound WIN",

abstract = "Rhinoviruses are the major causative agents of the common cold in humans. Here, we studied the stability of human rhinovirus type 14 (HRV14) under conditions of high hydrostatic pressure, low temperature, and urea in the absence and presence of an antiviral drug. Capsid dissociation and changes in the protein conformation were monitored by fluorescence spectroscopy, light scattering, circular dichroism, gel filtration chromatography, mass spectrometry and infectivity assays. The data show that high pressure induces the dissociation of HRV14 and that this process is inhibited by WIN 52084. MALDI-TOF mass spectrometry experiments demonstrate that VP4, the most internal viral protein, is released from the capsid by pressure treatment. This release of VP4 is concomitant with loss of infectivity. Our studies also show that at least one antiviral effect of the WIN drugs involves the locking of VP4 inside the capsid by blocking the dynamics associated with cell attachment.",

keywords = "WIN drugs, fluorescence, human rhinovirus, hydrostatic pressure, mass spectrometry",

author = "Gon{\c c}alves, {Rafael B.} and Mendes, {Ygara S.} and Soares, {Marcia R.} and Umesh Katpally and Smith, {Thomas J.} and Silva, {Jerson L.} and Oliveira, {Andr{\'e}a C.}",

note = "Funding Information: We are grateful to Rede Prote{\^o}mica do Estado do Rio de Janeiro for Mass Spectrometry facility and Emerson R. Gon{\c c}alves for competent technical assistance. We also thank Martha Sorenson for critical reading of the manuscript. This work was supported in part by a grant from the Millennium Institute Program (Millennium Institute for Structural Biology in Biomedicine and Biotechnology) to J.L.S. and A.C.O., by an international grant from the International Centre for Genetic Engineering and Biotechnology (ICGEB) to J.L.S., by an NIH grant to T.J.S. (GM10704) and by grants from Conselho Nacional de Desenvolvimento Cient{\'i}fico e Tecnol{\'o}gico (CNPq), Funda{\c c}{\~a}o Carlos Chagas Filho de Amparo {\`a} Pesquisa do Estado do Rio de Janeiro (FAPERJ), Funda{\c c}{\~a}o Universit{\'a}ria Jos{\'e} Bonif{\'a}cio (FUJB) and Financiadora de Estudos e Projetos (FINEP) to J.L.S. and A.C.O.",

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doi = "10.1016/j.jmb.2006.11.033",

language = "English (US)",

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T1 - VP4 Protein from Human Rhinovirus 14 Is Released by Pressure and Locked in the Capsid by the Antiviral Compound WIN

AU - Gonçalves, Rafael B.

AU - Mendes, Ygara S.

AU - Soares, Marcia R.

AU - Katpally, Umesh

AU - Smith, Thomas J.

AU - Silva, Jerson L.

AU - Oliveira, Andréa C.

N1 - Funding Information: We are grateful to Rede Proteômica do Estado do Rio de Janeiro for Mass Spectrometry facility and Emerson R. Gonçalves for competent technical assistance. We also thank Martha Sorenson for critical reading of the manuscript. This work was supported in part by a grant from the Millennium Institute Program (Millennium Institute for Structural Biology in Biomedicine and Biotechnology) to J.L.S. and A.C.O., by an international grant from the International Centre for Genetic Engineering and Biotechnology (ICGEB) to J.L.S., by an NIH grant to T.J.S. (GM10704) and by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Fundação Universitária José Bonifácio (FUJB) and Financiadora de Estudos e Projetos (FINEP) to J.L.S. and A.C.O.

PY - 2007/2/9

Y1 - 2007/2/9

N2 - Rhinoviruses are the major causative agents of the common cold in humans. Here, we studied the stability of human rhinovirus type 14 (HRV14) under conditions of high hydrostatic pressure, low temperature, and urea in the absence and presence of an antiviral drug. Capsid dissociation and changes in the protein conformation were monitored by fluorescence spectroscopy, light scattering, circular dichroism, gel filtration chromatography, mass spectrometry and infectivity assays. The data show that high pressure induces the dissociation of HRV14 and that this process is inhibited by WIN 52084. MALDI-TOF mass spectrometry experiments demonstrate that VP4, the most internal viral protein, is released from the capsid by pressure treatment. This release of VP4 is concomitant with loss of infectivity. Our studies also show that at least one antiviral effect of the WIN drugs involves the locking of VP4 inside the capsid by blocking the dynamics associated with cell attachment.

AB - Rhinoviruses are the major causative agents of the common cold in humans. Here, we studied the stability of human rhinovirus type 14 (HRV14) under conditions of high hydrostatic pressure, low temperature, and urea in the absence and presence of an antiviral drug. Capsid dissociation and changes in the protein conformation were monitored by fluorescence spectroscopy, light scattering, circular dichroism, gel filtration chromatography, mass spectrometry and infectivity assays. The data show that high pressure induces the dissociation of HRV14 and that this process is inhibited by WIN 52084. MALDI-TOF mass spectrometry experiments demonstrate that VP4, the most internal viral protein, is released from the capsid by pressure treatment. This release of VP4 is concomitant with loss of infectivity. Our studies also show that at least one antiviral effect of the WIN drugs involves the locking of VP4 inside the capsid by blocking the dynamics associated with cell attachment.

KW - WIN drugs

KW - fluorescence

KW - human rhinovirus

KW - hydrostatic pressure

KW - mass spectrometry

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JF - Journal of Molecular Biology

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ER -

VP4 Protein from Human Rhinovirus 14 Is Released by Pressure and Locked in the Capsid by the Antiviral Compound WIN

Abstract

Keywords

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