Isolation of Galectin-1 from Human Platelets: Its Interaction with Actin

Galectins are a family of animal lectins defined by their β-galactoside-binding specificity and a consensus sequence in their carbohydrate-recognition domain. Galectin-1 (Gal-1) is expressed as a non-covalently linked homodimer present in a variety of tissues. Here we describe its isolation from hum...

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Detalles Bibliográficos
Autores principales: González, Mariana Marta, Yoshizaki, L., Wolfenstein-Todel, C., Fink, Nilda Esther
Formato: Articulo
Lenguaje:Inglés
Publicado: 2012
Materias:
Ciencias Exactas
Biología
Actin
Aggregation
Galectin
Mass spectrometry
Platelets
Confocal microscopy
Hemagglutination
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/133565
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-133565
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Biología
Actin
Aggregation
Galectin
Mass spectrometry
Platelets
Confocal microscopy
Hemagglutination
spellingShingle Ciencias Exactas
Biología
Actin
Aggregation
Galectin
Mass spectrometry
Platelets
Confocal microscopy
Hemagglutination
González, Mariana Marta
Yoshizaki, L.
Wolfenstein-Todel, C.
Fink, Nilda Esther
Isolation of Galectin-1 from Human Platelets: Its Interaction with Actin
topic_facet Ciencias Exactas
Biología
Actin
Aggregation
Galectin
Mass spectrometry
Platelets
Confocal microscopy
Hemagglutination
description Galectins are a family of animal lectins defined by their β-galactoside-binding specificity and a consensus sequence in their carbohydrate-recognition domain. Galectin-1 (Gal-1) is expressed as a non-covalently linked homodimer present in a variety of tissues. Here we describe its isolation from human platelets by a procedure involving ionic exchange chromatography and affinity chromatography on lactose-agarose. Platelet Gal-1 co-purifies with actin, forming an actin-Gal-1 complex which does no dissociate even after treatment with sodium dodecyl sulfate. The presence of both proteins was confirmed by Western blot and by trypsin digestion followed by mass spectrometry identification. By hemagglutination assays we studied the response of recombinant Gal-1/actin, mixed and pre-incubated in different proportions, and then tested against neuraminidase treated rabbit red blood cells. The complex formation was confirmed by confocal microscopy, showing that both proteins co-localised in resting platelets as well as in thrombin-activated ones. These results suggest that endogenous Gal-1 forms an intracellular complex with monomeric actin and that, after platelet activation, Gal-1 could play a role in the polymerization-depolymerization process of actin, which concludes in platelet aggregation.
format Articulo
Articulo
author González, Mariana Marta
Yoshizaki, L.
Wolfenstein-Todel, C.
Fink, Nilda Esther
author_facet González, Mariana Marta
Yoshizaki, L.
Wolfenstein-Todel, C.
Fink, Nilda Esther
author_sort González, Mariana Marta
title Isolation of Galectin-1 from Human Platelets: Its Interaction with Actin
title_short Isolation of Galectin-1 from Human Platelets: Its Interaction with Actin
title_full Isolation of Galectin-1 from Human Platelets: Its Interaction with Actin
title_fullStr Isolation of Galectin-1 from Human Platelets: Its Interaction with Actin
title_full_unstemmed Isolation of Galectin-1 from Human Platelets: Its Interaction with Actin
title_sort isolation of galectin-1 from human platelets: its interaction with actin
publishDate 2012
url http://sedici.unlp.edu.ar/handle/10915/133565
work_keys_str_mv AT gonzalezmarianamarta isolationofgalectin1fromhumanplateletsitsinteractionwithactin
AT yoshizakil isolationofgalectin1fromhumanplateletsitsinteractionwithactin
AT wolfensteintodelc isolationofgalectin1fromhumanplateletsitsinteractionwithactin
AT finknildaesther isolationofgalectin1fromhumanplateletsitsinteractionwithactin
bdutipo_str Repositorios
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