Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles.

Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles. Research Abstract Details 

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Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles. Abstract Text:

Richard W Clarke,Nilah Monnier,Haitao Li,Dejian Zhou,Helena Browne,David Klenerman,Richard W Clarke,Nilah Monnier,Haitao Li,Dejian Zhou,Helena Browne,David Klenerman,

We present a single virion method to determine absolute distributions of copy number in the protein composition of viruses and apply it to herpes simplex virus type 1. Using two-color coincidence fluorescence spectroscopy, we determine the virion-to-virion variability in copy numbers of fluorescently labeled tegument and envelope proteins relative to a capsid protein by analyzing fluorescence intensity ratios for ensembles of individual dual-labeled virions and fitting the resulting histogram of ratios. Using EYFP-tagged capsid protein VP26 as a reference for fluorescence intensity, we are able to calculate the mean and also, for the first time to our knowledge, the variation in numbers of gD, VP16, and VP22 tegument. The measurement of the number of glycoprotein D molecules was in good agreement with independent measurements of average numbers of these glycoproteins in bulk virus preparations, validating the method. The accuracy, straightforward data processing, and high throughput of this technique make it widely applicable to the analysis of the molecular composition of large complexes in general, and it is particularly suited to providing insights into virus structure, assembly, and infectivity.

Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles. Publishing Authors By Initials

RW Clarke,N Monnier,H Li,D Zhou,H Browne,D Klenerman,RW Clarke,N Monnier,H Li,D Zhou,H Browne,D Klenerman,

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Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Biophysical journal

VOLUME: 93

Page Numbers: 1329-37

Journal Abbreviation: Biophys. J.

ISSN: 0006-3495

DAY: 18

MONTH: 05

YEAR: 2007

Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles. Information

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LANGUAGE: eng

NlmUniqueID: 370626

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Grant and Affiliation Information for Two-color fluorescence analysis of individual virions determines the distribution of the copy number of proteins in herpes simplex virus particles.

AFFILIATION: Department of Chemistry and Division of Virology, University of Cambridge, Cambridge, United Kingdom.

Country: United States

AGENCY: United Kingdom Wellcome T

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MEDLINETA: Biophys J

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