Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatography

The invasion of the erythrocyte by Plasmodium falciparum depends on the ability of the merozoite to move through the membrane invagination. This ability is probably mediated by actin dependent motors. Using affinity columns with G-actin and F-actin we isolated actin binding proteins from the parasit...

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Main Authors: Claudia Forero, Moisés Wasserman
Format: Article
Language:English
Published: Fundação Oswaldo Cruz (FIOCRUZ) 2000-06-01
Series:Memorias do Instituto Oswaldo Cruz
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762000000300007
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author Claudia Forero
Moisés Wasserman
author_facet Claudia Forero
Moisés Wasserman
author_sort Claudia Forero
collection DOAJ
description The invasion of the erythrocyte by Plasmodium falciparum depends on the ability of the merozoite to move through the membrane invagination. This ability is probably mediated by actin dependent motors. Using affinity columns with G-actin and F-actin we isolated actin binding proteins from the parasite. By immunoblotting and immunoprecipitation with specific antibodies we identified the presence of tropomyosin, myosin, a-actinin, and two different actins in the eluate corresponding to F-actin binding proteins. In addition to these, a 240-260 kDa doublet, different in size from the erythrocyte spectrin, reacted with an antibody against human spectrin. All the above mentioned proteins were metabolically radiolabeled when the parasite was cultured with 35S-methionine. The presence of these proteins in P. falciparum is indicative of a complex cytoskeleton and supports the proposed role for an actin-myosin motor during invasion.
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spelling doaj.art-ad133bb44e964c968b1436c848638cbd2023-08-02T09:32:15ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz0074-02761678-80602000-06-0195332933710.1590/S0074-02762000000300007Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatographyClaudia ForeroMoisés WassermanThe invasion of the erythrocyte by Plasmodium falciparum depends on the ability of the merozoite to move through the membrane invagination. This ability is probably mediated by actin dependent motors. Using affinity columns with G-actin and F-actin we isolated actin binding proteins from the parasite. By immunoblotting and immunoprecipitation with specific antibodies we identified the presence of tropomyosin, myosin, a-actinin, and two different actins in the eluate corresponding to F-actin binding proteins. In addition to these, a 240-260 kDa doublet, different in size from the erythrocyte spectrin, reacted with an antibody against human spectrin. All the above mentioned proteins were metabolically radiolabeled when the parasite was cultured with 35S-methionine. The presence of these proteins in P. falciparum is indicative of a complex cytoskeleton and supports the proposed role for an actin-myosin motor during invasion.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762000000300007actina-actininmalariamyosinspectrintropomyosinPlasmodium falciparumaffinity chromatography
spellingShingle Claudia Forero
Moisés Wasserman
Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatography
Memorias do Instituto Oswaldo Cruz
actin
a-actinin
malaria
myosin
spectrin
tropomyosin
Plasmodium falciparum
affinity chromatography
title Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatography
title_full Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatography
title_fullStr Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatography
title_full_unstemmed Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatography
title_short Isolation and identification of actin-binding proteins in Plasmodium falciparum by affinity chromatography
title_sort isolation and identification of actin binding proteins in plasmodium falciparum by affinity chromatography
topic actin
a-actinin
malaria
myosin
spectrin
tropomyosin
Plasmodium falciparum
affinity chromatography
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762000000300007
work_keys_str_mv AT claudiaforero isolationandidentificationofactinbindingproteinsinplasmodiumfalciparumbyaffinitychromatography
AT moiseswasserman isolationandidentificationofactinbindingproteinsinplasmodiumfalciparumbyaffinitychromatography