Calpain-dependent cleavage of cain/cabin1 activates calcineurin to mediate calcium-triggered cell death

Min Jung Kim, Dong Gyu Jo, Gil Sun Hong, Byung Ju Kim, Michael Lai, Dong Hyung Cho, Ki Woo Kim, Arun Bandyopadhyay, Yeon Mi Hong, Do Han Kim, Chunghee Cho, Jun O. Liu, Solomon H. Snyder, Yong Keun Jung

Research output: Contribution to journalArticle

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Abstract

Cain/cabin1 is an endogenous inhibitor of calcineurin (Cn), a calcium-dependent serine/threonine phosphatase involved in various cellular functions including apoptosis. We show here that during apoptosis cain/cabin1 is cleaved by calpain at the carboxyl terminus to generate a cleavage product with a molecular mass of 32 kDa as a necessary step leading to Cn-mediated cell death. Mouse cain/cabin1 was identified from a thymus cDNA library by an in vitro substrate-screening assay with calpain. Exposure of Jurkat cells to the calcium ionophore, A23187, induced cain/cabin1 cleavage and cell death, accompanied by activation of calpain and Cn. The calpain inhibitors, calpeptin and zLLY, suppressed both A23187-induced cain/cabin1 cleavage and Cn activation, indicating that Cn activation and cain/cabin1 cleavage are calpain-dependent. Expression of cain/cabin1 or a catalytically inactive Cn mutant [CnAβ2(1-401/H160N)] and treatment with FK506 reduced A23187-induced cell death. In vitro calpain cleavage and immuno-precipitation assays with deletion mutants of cain/cabin1 showed that cleavage occurred in the Cn-binding domain of cain/cabin1, indicating that the cleavage at its C terminus by calpain prevented cain/cabin1 from binding to Cn. In addition, in vitro binding assays showed that cain/cabin1 bound to the Cn B-binding domain of Cn A. Taken together, these results indicate that calpain cleaves the calcineurin-binding domain of cain/cabin1 to activate Cn and elicit calcium-triggered cell death.

Original languageEnglish
Pages (from-to)9870-9875
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume99
Issue number15
DOIs
Publication statusPublished - 2002 Jul 23

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Calpain
Calcineurin
Cell Death
Calcium
Calcimycin
Apoptosis
Jurkat Cells
Calcium Ionophores
Phosphoprotein Phosphatases
Tacrolimus
Gene Library
Thymus Gland

All Science Journal Classification (ASJC) codes

  • General

Cite this

Kim, Min Jung ; Jo, Dong Gyu ; Hong, Gil Sun ; Kim, Byung Ju ; Lai, Michael ; Cho, Dong Hyung ; Kim, Ki Woo ; Bandyopadhyay, Arun ; Hong, Yeon Mi ; Kim, Do Han ; Cho, Chunghee ; Liu, Jun O. ; Snyder, Solomon H. ; Jung, Yong Keun. / Calpain-dependent cleavage of cain/cabin1 activates calcineurin to mediate calcium-triggered cell death. In: Proceedings of the National Academy of Sciences of the United States of America. 2002 ; Vol. 99, No. 15. pp. 9870-9875.
@article{8e017e2cb10b48b1a555a081a99b8728,
title = "Calpain-dependent cleavage of cain/cabin1 activates calcineurin to mediate calcium-triggered cell death",
abstract = "Cain/cabin1 is an endogenous inhibitor of calcineurin (Cn), a calcium-dependent serine/threonine phosphatase involved in various cellular functions including apoptosis. We show here that during apoptosis cain/cabin1 is cleaved by calpain at the carboxyl terminus to generate a cleavage product with a molecular mass of 32 kDa as a necessary step leading to Cn-mediated cell death. Mouse cain/cabin1 was identified from a thymus cDNA library by an in vitro substrate-screening assay with calpain. Exposure of Jurkat cells to the calcium ionophore, A23187, induced cain/cabin1 cleavage and cell death, accompanied by activation of calpain and Cn. The calpain inhibitors, calpeptin and zLLY, suppressed both A23187-induced cain/cabin1 cleavage and Cn activation, indicating that Cn activation and cain/cabin1 cleavage are calpain-dependent. Expression of cain/cabin1 or a catalytically inactive Cn mutant [CnAβ2(1-401/H160N)] and treatment with FK506 reduced A23187-induced cell death. In vitro calpain cleavage and immuno-precipitation assays with deletion mutants of cain/cabin1 showed that cleavage occurred in the Cn-binding domain of cain/cabin1, indicating that the cleavage at its C terminus by calpain prevented cain/cabin1 from binding to Cn. In addition, in vitro binding assays showed that cain/cabin1 bound to the Cn B-binding domain of Cn A. Taken together, these results indicate that calpain cleaves the calcineurin-binding domain of cain/cabin1 to activate Cn and elicit calcium-triggered cell death.",
author = "Kim, {Min Jung} and Jo, {Dong Gyu} and Hong, {Gil Sun} and Kim, {Byung Ju} and Michael Lai and Cho, {Dong Hyung} and Kim, {Ki Woo} and Arun Bandyopadhyay and Hong, {Yeon Mi} and Kim, {Do Han} and Chunghee Cho and Liu, {Jun O.} and Snyder, {Solomon H.} and Jung, {Yong Keun}",
year = "2002",
month = "7",
day = "23",
doi = "10.1073/pnas.152336999",
language = "English",
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Kim, MJ, Jo, DG, Hong, GS, Kim, BJ, Lai, M, Cho, DH, Kim, KW, Bandyopadhyay, A, Hong, YM, Kim, DH, Cho, C, Liu, JO, Snyder, SH & Jung, YK 2002, 'Calpain-dependent cleavage of cain/cabin1 activates calcineurin to mediate calcium-triggered cell death', Proceedings of the National Academy of Sciences of the United States of America, vol. 99, no. 15, pp. 9870-9875. https://doi.org/10.1073/pnas.152336999

Calpain-dependent cleavage of cain/cabin1 activates calcineurin to mediate calcium-triggered cell death. / Kim, Min Jung; Jo, Dong Gyu; Hong, Gil Sun; Kim, Byung Ju; Lai, Michael; Cho, Dong Hyung; Kim, Ki Woo; Bandyopadhyay, Arun; Hong, Yeon Mi; Kim, Do Han; Cho, Chunghee; Liu, Jun O.; Snyder, Solomon H.; Jung, Yong Keun.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, No. 15, 23.07.2002, p. 9870-9875.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Calpain-dependent cleavage of cain/cabin1 activates calcineurin to mediate calcium-triggered cell death

AU - Kim, Min Jung

AU - Jo, Dong Gyu

AU - Hong, Gil Sun

AU - Kim, Byung Ju

AU - Lai, Michael

AU - Cho, Dong Hyung

AU - Kim, Ki Woo

AU - Bandyopadhyay, Arun

AU - Hong, Yeon Mi

AU - Kim, Do Han

AU - Cho, Chunghee

AU - Liu, Jun O.

AU - Snyder, Solomon H.

AU - Jung, Yong Keun

PY - 2002/7/23

Y1 - 2002/7/23

N2 - Cain/cabin1 is an endogenous inhibitor of calcineurin (Cn), a calcium-dependent serine/threonine phosphatase involved in various cellular functions including apoptosis. We show here that during apoptosis cain/cabin1 is cleaved by calpain at the carboxyl terminus to generate a cleavage product with a molecular mass of 32 kDa as a necessary step leading to Cn-mediated cell death. Mouse cain/cabin1 was identified from a thymus cDNA library by an in vitro substrate-screening assay with calpain. Exposure of Jurkat cells to the calcium ionophore, A23187, induced cain/cabin1 cleavage and cell death, accompanied by activation of calpain and Cn. The calpain inhibitors, calpeptin and zLLY, suppressed both A23187-induced cain/cabin1 cleavage and Cn activation, indicating that Cn activation and cain/cabin1 cleavage are calpain-dependent. Expression of cain/cabin1 or a catalytically inactive Cn mutant [CnAβ2(1-401/H160N)] and treatment with FK506 reduced A23187-induced cell death. In vitro calpain cleavage and immuno-precipitation assays with deletion mutants of cain/cabin1 showed that cleavage occurred in the Cn-binding domain of cain/cabin1, indicating that the cleavage at its C terminus by calpain prevented cain/cabin1 from binding to Cn. In addition, in vitro binding assays showed that cain/cabin1 bound to the Cn B-binding domain of Cn A. Taken together, these results indicate that calpain cleaves the calcineurin-binding domain of cain/cabin1 to activate Cn and elicit calcium-triggered cell death.

AB - Cain/cabin1 is an endogenous inhibitor of calcineurin (Cn), a calcium-dependent serine/threonine phosphatase involved in various cellular functions including apoptosis. We show here that during apoptosis cain/cabin1 is cleaved by calpain at the carboxyl terminus to generate a cleavage product with a molecular mass of 32 kDa as a necessary step leading to Cn-mediated cell death. Mouse cain/cabin1 was identified from a thymus cDNA library by an in vitro substrate-screening assay with calpain. Exposure of Jurkat cells to the calcium ionophore, A23187, induced cain/cabin1 cleavage and cell death, accompanied by activation of calpain and Cn. The calpain inhibitors, calpeptin and zLLY, suppressed both A23187-induced cain/cabin1 cleavage and Cn activation, indicating that Cn activation and cain/cabin1 cleavage are calpain-dependent. Expression of cain/cabin1 or a catalytically inactive Cn mutant [CnAβ2(1-401/H160N)] and treatment with FK506 reduced A23187-induced cell death. In vitro calpain cleavage and immuno-precipitation assays with deletion mutants of cain/cabin1 showed that cleavage occurred in the Cn-binding domain of cain/cabin1, indicating that the cleavage at its C terminus by calpain prevented cain/cabin1 from binding to Cn. In addition, in vitro binding assays showed that cain/cabin1 bound to the Cn B-binding domain of Cn A. Taken together, these results indicate that calpain cleaves the calcineurin-binding domain of cain/cabin1 to activate Cn and elicit calcium-triggered cell death.

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JF - Proceedings of the National Academy of Sciences of the United States of America

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