Concanavalin A aggregation and toxicity on cell cultures

Valeria Militello, Valeria Vetri, Pasquale Picone, Rita Carrotta, Marta Di Carlo

Risultato della ricerca: Article

28 Citazioni (Scopus)

Abstract

A number of neurodegenerative diseases are known to involve protein aggregation. Common mechanisms and structural properties of amyloids are thought to be involved in aggregation-related cytotoxicity. In this context we propose an experimental study on Concanavalin A (Con A) aggregation and use it as a model to study the relationship between cell toxicity and aggregation processes. Depending on solution conditions, Con A aggregation has been monitored by static and dynamic light scattering, Thioflavin T emission, and FTIR absorption. The morphology of different aggregate species was verified by means of Atomic Force Microscopy and Confocal Microscopy. During the aggregation pathway the native protein conformation is destabilized and as a consequence, the simultaneous occurrence of conformational changes and protein aggregation is observed in both conditions. The effects of the extracellular addition of native protein, oligomers and mature fibrils were tested on LAN5 neuroblastoma cells by MTS assay. Results showed the toxicity of the first two species while a negligible effect was detected for amyloid fibrils. Both native and oligomeric aggregates were found to be able to activate apoptosis exclusively by extrinsic pathway through caspase 8 activation. Those results suggest that cytotoxicity mechanisms arise from specific membrane interactions with reactive conformations of destabilized molecules occurring during the amyloidal aggregation pathway. Those conformations, populated when native or preformed oligomers are incubated, are unavailable to bind cell membrane proteins. This happens because they are recruited in the mature fibrillar structure which-as a consequence-turns out to be non-toxic.
Lingua originaleEnglish
pagine (da-a)173-183
Numero di pagine11
RivistaBIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS
Volume1804
Stato di pubblicazionePublished - 2010

Fingerprint

Concanavalin A
Cell culture
Toxicity
Agglomeration
Cell Culture Techniques
Amyloid
Cell Aggregation
Protein Conformation
Proteins
Caspase 8
Atomic Force Microscopy
Fourier Transform Infrared Spectroscopy
Neuroblastoma
Conformations
Confocal Microscopy
Neurodegenerative Diseases
Membrane Proteins
Cytotoxicity
Oligomers
Apoptosis

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biophysics
  • Biochemistry
  • Molecular Biology

Cita questo

Concanavalin A aggregation and toxicity on cell cultures. / Militello, Valeria; Vetri, Valeria; Picone, Pasquale; Carrotta, Rita; Di Carlo, Marta.

In: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, Vol. 1804, 2010, pag. 173-183.

Risultato della ricerca: Article

Militello, Valeria ; Vetri, Valeria ; Picone, Pasquale ; Carrotta, Rita ; Di Carlo, Marta. / Concanavalin A aggregation and toxicity on cell cultures. In: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS. 2010 ; Vol. 1804. pagg. 173-183.
@article{60251cad92f94d768705f18fd35d2f54,
title = "Concanavalin A aggregation and toxicity on cell cultures",
abstract = "A number of neurodegenerative diseases are known to involve protein aggregation. Common mechanisms and structural properties of amyloids are thought to be involved in aggregation-related cytotoxicity. In this context we propose an experimental study on Concanavalin A (Con A) aggregation and use it as a model to study the relationship between cell toxicity and aggregation processes. Depending on solution conditions, Con A aggregation has been monitored by static and dynamic light scattering, Thioflavin T emission, and FTIR absorption. The morphology of different aggregate species was verified by means of Atomic Force Microscopy and Confocal Microscopy. During the aggregation pathway the native protein conformation is destabilized and as a consequence, the simultaneous occurrence of conformational changes and protein aggregation is observed in both conditions. The effects of the extracellular addition of native protein, oligomers and mature fibrils were tested on LAN5 neuroblastoma cells by MTS assay. Results showed the toxicity of the first two species while a negligible effect was detected for amyloid fibrils. Both native and oligomeric aggregates were found to be able to activate apoptosis exclusively by extrinsic pathway through caspase 8 activation. Those results suggest that cytotoxicity mechanisms arise from specific membrane interactions with reactive conformations of destabilized molecules occurring during the amyloidal aggregation pathway. Those conformations, populated when native or preformed oligomers are incubated, are unavailable to bind cell membrane proteins. This happens because they are recruited in the mature fibrillar structure which-as a consequence-turns out to be non-toxic.",
keywords = "Amyloids, Citotoxicity, Oligomers, Protein aggregation",
author = "Valeria Militello and Valeria Vetri and Pasquale Picone and Rita Carrotta and {Di Carlo}, Marta",
year = "2010",
language = "English",
volume = "1804",
pages = "173--183",
journal = "Biochimica et Biophysica Acta - Proteins and Proteomics",
issn = "1570-9639",
publisher = "Elsevier",

}

TY - JOUR

T1 - Concanavalin A aggregation and toxicity on cell cultures

AU - Militello, Valeria

AU - Vetri, Valeria

AU - Picone, Pasquale

AU - Carrotta, Rita

AU - Di Carlo, Marta

PY - 2010

Y1 - 2010

N2 - A number of neurodegenerative diseases are known to involve protein aggregation. Common mechanisms and structural properties of amyloids are thought to be involved in aggregation-related cytotoxicity. In this context we propose an experimental study on Concanavalin A (Con A) aggregation and use it as a model to study the relationship between cell toxicity and aggregation processes. Depending on solution conditions, Con A aggregation has been monitored by static and dynamic light scattering, Thioflavin T emission, and FTIR absorption. The morphology of different aggregate species was verified by means of Atomic Force Microscopy and Confocal Microscopy. During the aggregation pathway the native protein conformation is destabilized and as a consequence, the simultaneous occurrence of conformational changes and protein aggregation is observed in both conditions. The effects of the extracellular addition of native protein, oligomers and mature fibrils were tested on LAN5 neuroblastoma cells by MTS assay. Results showed the toxicity of the first two species while a negligible effect was detected for amyloid fibrils. Both native and oligomeric aggregates were found to be able to activate apoptosis exclusively by extrinsic pathway through caspase 8 activation. Those results suggest that cytotoxicity mechanisms arise from specific membrane interactions with reactive conformations of destabilized molecules occurring during the amyloidal aggregation pathway. Those conformations, populated when native or preformed oligomers are incubated, are unavailable to bind cell membrane proteins. This happens because they are recruited in the mature fibrillar structure which-as a consequence-turns out to be non-toxic.

AB - A number of neurodegenerative diseases are known to involve protein aggregation. Common mechanisms and structural properties of amyloids are thought to be involved in aggregation-related cytotoxicity. In this context we propose an experimental study on Concanavalin A (Con A) aggregation and use it as a model to study the relationship between cell toxicity and aggregation processes. Depending on solution conditions, Con A aggregation has been monitored by static and dynamic light scattering, Thioflavin T emission, and FTIR absorption. The morphology of different aggregate species was verified by means of Atomic Force Microscopy and Confocal Microscopy. During the aggregation pathway the native protein conformation is destabilized and as a consequence, the simultaneous occurrence of conformational changes and protein aggregation is observed in both conditions. The effects of the extracellular addition of native protein, oligomers and mature fibrils were tested on LAN5 neuroblastoma cells by MTS assay. Results showed the toxicity of the first two species while a negligible effect was detected for amyloid fibrils. Both native and oligomeric aggregates were found to be able to activate apoptosis exclusively by extrinsic pathway through caspase 8 activation. Those results suggest that cytotoxicity mechanisms arise from specific membrane interactions with reactive conformations of destabilized molecules occurring during the amyloidal aggregation pathway. Those conformations, populated when native or preformed oligomers are incubated, are unavailable to bind cell membrane proteins. This happens because they are recruited in the mature fibrillar structure which-as a consequence-turns out to be non-toxic.

KW - Amyloids

KW - Citotoxicity

KW - Oligomers

KW - Protein aggregation

UR - http://hdl.handle.net/10447/46176

UR - http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B73DJ-4X9NV7P-1&_user=519924&_coverDate=01%2F31%2F2010&_rdoc=1&_fmt=high&_orig=search&_sort=d&_docanchor=&view=c&_acct=C000025965&_version=1&_urlVersion=0&_userid=519924&md5=8b341bd7079569049b386a163ee75818

M3 - Article

VL - 1804

SP - 173

EP - 183

JO - Biochimica et Biophysica Acta - Proteins and Proteomics

JF - Biochimica et Biophysica Acta - Proteins and Proteomics

SN - 1570-9639

ER -