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High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar

The structural polypeptides of baculoviruses of Autographa californica (AcMNPV) and Porthetria dispar (PdMNPV) were analyzed by two-dimensional (2-D) gel electrophoresis. Purified proteins were solubilized in urea-NP40 mix and separated by isoelectric focusing in the first dimension; electrophoresis...

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Autores principales: Singh, Shiva P., Gudauskas, Robert T., Harper, James D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Published by Elsevier Inc. 1983
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7130522/
https://www.ncbi.nlm.nih.gov/pubmed/18638911
http://dx.doi.org/10.1016/0042-6822(83)90209-X
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author Singh, Shiva P.
Gudauskas, Robert T.
Harper, James D.
author_facet Singh, Shiva P.
Gudauskas, Robert T.
Harper, James D.
author_sort Singh, Shiva P.
collection PubMed
description The structural polypeptides of baculoviruses of Autographa californica (AcMNPV) and Porthetria dispar (PdMNPV) were analyzed by two-dimensional (2-D) gel electrophoresis. Purified proteins were solubilized in urea-NP40 mix and separated by isoelectric focusing in the first dimension; electrophoresis in the presence of sodium dodecyl sulfate (SDS) separated proteins by molecular weight in the second dimension. Eighty-one acidic polypeptides ranging in molecular weight from 13,500 to 86,000 Da were resolved in AcMNPV enveloped virions. The predominant polypeptide had a molecular weight of 41,500 and was considered to be the major capsid protein. Nucleocapsids from AcMNPV were resolved into 64 polypeptides. At least 11 of the polypeptides, including most of the high molecular weight proteins, that were not resolved in nucleocapsids were considered to be envelope proteins. For PdMNPV enveloped virions, there were 95 acidic polypeptides ranging in molecular weight from 13,500 to 85,500. The predominant polypeptide had a molecular weight of 46,500 Da. Polyhedral proteins (polyhedrin) isolated from protease-inactivated polyhedra and separable into a single major polypeptide (approx. 31,000) on one-dimensional SDS-polyacrylamide gel electrophoresis were resolved into six polypeptides for both viruses. All six polyhedrin polypeptides had the same molecular weight, but their isoelectric points ranged from pH 5.3 to 5.9 for AcMNPV and from pH 5.7 to 6.2 for PdMNPV. These six polypeptides were also detected when protease-inactivated or noninactivated whole polyhedra were analyzed directly by 2-D electrophoresis. It is assumed that not all the observed baculovirus polypeptides were unique species. Some proteins, especially the polyhedrin polypeptides, appeared to be related and had altered mobilities as a consequence of post-translational modifications.
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spelling pubmed-71305222020-04-08 High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar Singh, Shiva P. Gudauskas, Robert T. Harper, James D. Virology Article The structural polypeptides of baculoviruses of Autographa californica (AcMNPV) and Porthetria dispar (PdMNPV) were analyzed by two-dimensional (2-D) gel electrophoresis. Purified proteins were solubilized in urea-NP40 mix and separated by isoelectric focusing in the first dimension; electrophoresis in the presence of sodium dodecyl sulfate (SDS) separated proteins by molecular weight in the second dimension. Eighty-one acidic polypeptides ranging in molecular weight from 13,500 to 86,000 Da were resolved in AcMNPV enveloped virions. The predominant polypeptide had a molecular weight of 41,500 and was considered to be the major capsid protein. Nucleocapsids from AcMNPV were resolved into 64 polypeptides. At least 11 of the polypeptides, including most of the high molecular weight proteins, that were not resolved in nucleocapsids were considered to be envelope proteins. For PdMNPV enveloped virions, there were 95 acidic polypeptides ranging in molecular weight from 13,500 to 85,500. The predominant polypeptide had a molecular weight of 46,500 Da. Polyhedral proteins (polyhedrin) isolated from protease-inactivated polyhedra and separable into a single major polypeptide (approx. 31,000) on one-dimensional SDS-polyacrylamide gel electrophoresis were resolved into six polypeptides for both viruses. All six polyhedrin polypeptides had the same molecular weight, but their isoelectric points ranged from pH 5.3 to 5.9 for AcMNPV and from pH 5.7 to 6.2 for PdMNPV. These six polypeptides were also detected when protease-inactivated or noninactivated whole polyhedra were analyzed directly by 2-D electrophoresis. It is assumed that not all the observed baculovirus polypeptides were unique species. Some proteins, especially the polyhedrin polypeptides, appeared to be related and had altered mobilities as a consequence of post-translational modifications. Published by Elsevier Inc. 1983-03 2004-02-06 /pmc/articles/PMC7130522/ /pubmed/18638911 http://dx.doi.org/10.1016/0042-6822(83)90209-X Text en Copyright © 1983 Published by Elsevier Inc. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Singh, Shiva P.
Gudauskas, Robert T.
Harper, James D.
High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar
title High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar
title_full High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar
title_fullStr High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar
title_full_unstemmed High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar
title_short High resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of Autographa californica and Porthetria (lymantria) dispar
title_sort high resolution two-dimensional gel electrophoresis of structural proteins of baculoviruses of autographa californica and porthetria (lymantria) dispar
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7130522/
https://www.ncbi.nlm.nih.gov/pubmed/18638911
http://dx.doi.org/10.1016/0042-6822(83)90209-X
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