A quantitative method for the specific assessment of caspase-6 activity in cell culture

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Standard

A quantitative method for the specific assessment of caspase-6 activity in cell culture. / Ehrnhoefer, Dagmar E; Skotte, Niels H; Savill, Jane; Nguyen, Yen T N; Ladha, Safia; Cao, Li-Ping; Dullaghan, Edie; Hayden, Michael R.

In: PLOS ONE, Vol. 6, No. 11, 2011, p. e27680.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Ehrnhoefer, DE, Skotte, NH, Savill, J, Nguyen, YTN, Ladha, S, Cao, L-P, Dullaghan, E & Hayden, MR 2011, 'A quantitative method for the specific assessment of caspase-6 activity in cell culture', PLOS ONE, vol. 6, no. 11, pp. e27680. https://doi.org/10.1371/journal.pone.0027680

APA

Ehrnhoefer, D. E., Skotte, N. H., Savill, J., Nguyen, Y. T. N., Ladha, S., Cao, L-P., Dullaghan, E., & Hayden, M. R. (2011). A quantitative method for the specific assessment of caspase-6 activity in cell culture. PLOS ONE, 6(11), e27680. https://doi.org/10.1371/journal.pone.0027680

Vancouver

Ehrnhoefer DE, Skotte NH, Savill J, Nguyen YTN, Ladha S, Cao L-P et al. A quantitative method for the specific assessment of caspase-6 activity in cell culture. PLOS ONE. 2011;6(11):e27680. https://doi.org/10.1371/journal.pone.0027680

Author

Ehrnhoefer, Dagmar E ; Skotte, Niels H ; Savill, Jane ; Nguyen, Yen T N ; Ladha, Safia ; Cao, Li-Ping ; Dullaghan, Edie ; Hayden, Michael R. / A quantitative method for the specific assessment of caspase-6 activity in cell culture. In: PLOS ONE. 2011 ; Vol. 6, No. 11. pp. e27680.

Bibtex

@article{ba0a0c8f5c8e41f8a49f0986b60ecef8,
title = "A quantitative method for the specific assessment of caspase-6 activity in cell culture",
abstract = "Aberrant activation of caspase-6 has recently emerged as a major contributor to the pathogeneses of neurodegenerative disorders such as Alzheimer's and Huntington disease. Commercially available assays to measure caspase-6 activity commonly use the VEID peptide as a substrate. However these methods are not well suited to specifically assess caspase-6 activity in the presence of other, confounding protease activities, as often encountered in cell and tissue samples. Here we report the development of a method that overcomes this limitation by using a protein substrate, lamin A, which is highly specific for caspase-6 cleavage at amino acid 230. Using a neo-epitope antibody against cleaved lamin A, we developed an electrochemiluminescence-based ELISA assay that is suitable to specifically detect and quantify caspase-6 activity in highly apoptotic cell extracts. The method is more sensitive than VEID-based assays and can be adapted to a high-content imaging platform for high-throughput screening. This method should be useful to screen for and characterize caspase-6 inhibitor compounds and other interventions to decrease intracellular caspase-6 activity for applications in neurodegenerative disorders.",
keywords = "Amino Acid Sequence, Animals, COS Cells, Caspase 6, Cell Culture Techniques, Cell Extracts, Cercopithecus aethiops, Enzyme Assays, Enzyme-Linked Immunosorbent Assay, Fluorescent Antibody Technique, Humans, Kinetics, Lamins, Luminescence, Mice, Molecular Sequence Data, Neurons, Peptides, Substrate Specificity",
author = "Ehrnhoefer, {Dagmar E} and Skotte, {Niels H} and Jane Savill and Nguyen, {Yen T N} and Safia Ladha and Li-Ping Cao and Edie Dullaghan and Hayden, {Michael R}",
year = "2011",
doi = "10.1371/journal.pone.0027680",
language = "English",
volume = "6",
pages = "e27680",
journal = "PLoS ONE",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "11",

}

RIS

TY - JOUR

T1 - A quantitative method for the specific assessment of caspase-6 activity in cell culture

AU - Ehrnhoefer, Dagmar E

AU - Skotte, Niels H

AU - Savill, Jane

AU - Nguyen, Yen T N

AU - Ladha, Safia

AU - Cao, Li-Ping

AU - Dullaghan, Edie

AU - Hayden, Michael R

PY - 2011

Y1 - 2011

N2 - Aberrant activation of caspase-6 has recently emerged as a major contributor to the pathogeneses of neurodegenerative disorders such as Alzheimer's and Huntington disease. Commercially available assays to measure caspase-6 activity commonly use the VEID peptide as a substrate. However these methods are not well suited to specifically assess caspase-6 activity in the presence of other, confounding protease activities, as often encountered in cell and tissue samples. Here we report the development of a method that overcomes this limitation by using a protein substrate, lamin A, which is highly specific for caspase-6 cleavage at amino acid 230. Using a neo-epitope antibody against cleaved lamin A, we developed an electrochemiluminescence-based ELISA assay that is suitable to specifically detect and quantify caspase-6 activity in highly apoptotic cell extracts. The method is more sensitive than VEID-based assays and can be adapted to a high-content imaging platform for high-throughput screening. This method should be useful to screen for and characterize caspase-6 inhibitor compounds and other interventions to decrease intracellular caspase-6 activity for applications in neurodegenerative disorders.

AB - Aberrant activation of caspase-6 has recently emerged as a major contributor to the pathogeneses of neurodegenerative disorders such as Alzheimer's and Huntington disease. Commercially available assays to measure caspase-6 activity commonly use the VEID peptide as a substrate. However these methods are not well suited to specifically assess caspase-6 activity in the presence of other, confounding protease activities, as often encountered in cell and tissue samples. Here we report the development of a method that overcomes this limitation by using a protein substrate, lamin A, which is highly specific for caspase-6 cleavage at amino acid 230. Using a neo-epitope antibody against cleaved lamin A, we developed an electrochemiluminescence-based ELISA assay that is suitable to specifically detect and quantify caspase-6 activity in highly apoptotic cell extracts. The method is more sensitive than VEID-based assays and can be adapted to a high-content imaging platform for high-throughput screening. This method should be useful to screen for and characterize caspase-6 inhibitor compounds and other interventions to decrease intracellular caspase-6 activity for applications in neurodegenerative disorders.

KW - Amino Acid Sequence

KW - Animals

KW - COS Cells

KW - Caspase 6

KW - Cell Culture Techniques

KW - Cell Extracts

KW - Cercopithecus aethiops

KW - Enzyme Assays

KW - Enzyme-Linked Immunosorbent Assay

KW - Fluorescent Antibody Technique

KW - Humans

KW - Kinetics

KW - Lamins

KW - Luminescence

KW - Mice

KW - Molecular Sequence Data

KW - Neurons

KW - Peptides

KW - Substrate Specificity

U2 - 10.1371/journal.pone.0027680

DO - 10.1371/journal.pone.0027680

M3 - Journal article

C2 - 22140457

VL - 6

SP - e27680

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 11

ER -

ID: 153451326