The caspase-3 cleavage product of the plasma membrane Ca 2+-ATPase 4b is activated and appropriately targeted

K. Pászty, Géza Antalffy, Alan R. Penheiter, L. Homolya, Rita Padányi, Attila Iliás, Adelaida G. Filoteo, John T. Penniston, A. Enyedi

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The calmodulin-activated transporter hPMCA4 (human plasma membrane Ca 2+-ATPase isoform 4) is a target for cleavage by caspase-3 during apoptosis. We have demonstrated that caspase-3 generates a 120 kDa fragment of this pump which lacks the complete autoinhibitory sequence [Paszty, Verma, Padanyi, Filoteo, Penniston and Enyedi (2002) J. Biol. Chem. 277, 6822-6829]. In the present study we analysed further the characteristics of the fragment of hPMCA4b produced by caspase-3. We did this by overexpressing the caspase-3 cleavage product of hPMCA4b in COS-7 and MDCKII (Madin-Darby canine kidney II) cells. This technique made it possible to clearly define the properties of this fragment, and we showed that it is constitutively active, as it forms a phosphoenzyme intermediate and has high Ca2+ transport activity in the absence of calmodulin. When this fragment of hPMCA4b was stably expressed in MDCKII cell clones, it was targeted without degradation to the basolateral plasma membrane. In summary, our studies emphasize that the caspase-3 cleavage product of hPMCA4b is constitutively active, and that the C-terminus is not required for proper targeting of hPMCA4b to the plasma membrane. Also, for the first time, we have generated cell clones that stably express a constitutively active PMCA.

Original languageEnglish
Pages (from-to)687-692
Number of pages6
JournalBiochemical Journal
Volume391
Issue number3
DOIs
Publication statusPublished - Nov 1 2005

Fingerprint

Cell membranes
Caspase 3
Adenosine Triphosphatases
Cell Membrane
Clone cells
Madin Darby Canine Kidney Cells
Calmodulin
Clone Cells
Protein Isoforms
Pumps
Apoptosis
Degradation

Keywords

  • Activation
  • Apoptosis
  • Calmodulin
  • Caspase-3
  • Localization
  • Plasma membrane Ca-ATPase

ASJC Scopus subject areas

  • Biochemistry

Cite this

The caspase-3 cleavage product of the plasma membrane Ca 2+-ATPase 4b is activated and appropriately targeted. / Pászty, K.; Antalffy, Géza; Penheiter, Alan R.; Homolya, L.; Padányi, Rita; Iliás, Attila; Filoteo, Adelaida G.; Penniston, John T.; Enyedi, A.

In: Biochemical Journal, Vol. 391, No. 3, 01.11.2005, p. 687-692.

Research output: Contribution to journalArticle

Pászty, K. ; Antalffy, Géza ; Penheiter, Alan R. ; Homolya, L. ; Padányi, Rita ; Iliás, Attila ; Filoteo, Adelaida G. ; Penniston, John T. ; Enyedi, A. / The caspase-3 cleavage product of the plasma membrane Ca 2+-ATPase 4b is activated and appropriately targeted. In: Biochemical Journal. 2005 ; Vol. 391, No. 3. pp. 687-692.
@article{8444cc4ebf204378adbba80bb97a72cf,
title = "The caspase-3 cleavage product of the plasma membrane Ca 2+-ATPase 4b is activated and appropriately targeted",
abstract = "The calmodulin-activated transporter hPMCA4 (human plasma membrane Ca 2+-ATPase isoform 4) is a target for cleavage by caspase-3 during apoptosis. We have demonstrated that caspase-3 generates a 120 kDa fragment of this pump which lacks the complete autoinhibitory sequence [Paszty, Verma, Padanyi, Filoteo, Penniston and Enyedi (2002) J. Biol. Chem. 277, 6822-6829]. In the present study we analysed further the characteristics of the fragment of hPMCA4b produced by caspase-3. We did this by overexpressing the caspase-3 cleavage product of hPMCA4b in COS-7 and MDCKII (Madin-Darby canine kidney II) cells. This technique made it possible to clearly define the properties of this fragment, and we showed that it is constitutively active, as it forms a phosphoenzyme intermediate and has high Ca2+ transport activity in the absence of calmodulin. When this fragment of hPMCA4b was stably expressed in MDCKII cell clones, it was targeted without degradation to the basolateral plasma membrane. In summary, our studies emphasize that the caspase-3 cleavage product of hPMCA4b is constitutively active, and that the C-terminus is not required for proper targeting of hPMCA4b to the plasma membrane. Also, for the first time, we have generated cell clones that stably express a constitutively active PMCA.",
keywords = "Activation, Apoptosis, Calmodulin, Caspase-3, Localization, Plasma membrane Ca-ATPase",
author = "K. P{\'a}szty and G{\'e}za Antalffy and Penheiter, {Alan R.} and L. Homolya and Rita Pad{\'a}nyi and Attila Ili{\'a}s and Filoteo, {Adelaida G.} and Penniston, {John T.} and A. Enyedi",
year = "2005",
month = "11",
day = "1",
doi = "10.1042/BJ20051012",
language = "English",
volume = "391",
pages = "687--692",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "3",

}

TY - JOUR

T1 - The caspase-3 cleavage product of the plasma membrane Ca 2+-ATPase 4b is activated and appropriately targeted

AU - Pászty, K.

AU - Antalffy, Géza

AU - Penheiter, Alan R.

AU - Homolya, L.

AU - Padányi, Rita

AU - Iliás, Attila

AU - Filoteo, Adelaida G.

AU - Penniston, John T.

AU - Enyedi, A.

PY - 2005/11/1

Y1 - 2005/11/1

N2 - The calmodulin-activated transporter hPMCA4 (human plasma membrane Ca 2+-ATPase isoform 4) is a target for cleavage by caspase-3 during apoptosis. We have demonstrated that caspase-3 generates a 120 kDa fragment of this pump which lacks the complete autoinhibitory sequence [Paszty, Verma, Padanyi, Filoteo, Penniston and Enyedi (2002) J. Biol. Chem. 277, 6822-6829]. In the present study we analysed further the characteristics of the fragment of hPMCA4b produced by caspase-3. We did this by overexpressing the caspase-3 cleavage product of hPMCA4b in COS-7 and MDCKII (Madin-Darby canine kidney II) cells. This technique made it possible to clearly define the properties of this fragment, and we showed that it is constitutively active, as it forms a phosphoenzyme intermediate and has high Ca2+ transport activity in the absence of calmodulin. When this fragment of hPMCA4b was stably expressed in MDCKII cell clones, it was targeted without degradation to the basolateral plasma membrane. In summary, our studies emphasize that the caspase-3 cleavage product of hPMCA4b is constitutively active, and that the C-terminus is not required for proper targeting of hPMCA4b to the plasma membrane. Also, for the first time, we have generated cell clones that stably express a constitutively active PMCA.

AB - The calmodulin-activated transporter hPMCA4 (human plasma membrane Ca 2+-ATPase isoform 4) is a target for cleavage by caspase-3 during apoptosis. We have demonstrated that caspase-3 generates a 120 kDa fragment of this pump which lacks the complete autoinhibitory sequence [Paszty, Verma, Padanyi, Filoteo, Penniston and Enyedi (2002) J. Biol. Chem. 277, 6822-6829]. In the present study we analysed further the characteristics of the fragment of hPMCA4b produced by caspase-3. We did this by overexpressing the caspase-3 cleavage product of hPMCA4b in COS-7 and MDCKII (Madin-Darby canine kidney II) cells. This technique made it possible to clearly define the properties of this fragment, and we showed that it is constitutively active, as it forms a phosphoenzyme intermediate and has high Ca2+ transport activity in the absence of calmodulin. When this fragment of hPMCA4b was stably expressed in MDCKII cell clones, it was targeted without degradation to the basolateral plasma membrane. In summary, our studies emphasize that the caspase-3 cleavage product of hPMCA4b is constitutively active, and that the C-terminus is not required for proper targeting of hPMCA4b to the plasma membrane. Also, for the first time, we have generated cell clones that stably express a constitutively active PMCA.

KW - Activation

KW - Apoptosis

KW - Calmodulin

KW - Caspase-3

KW - Localization

KW - Plasma membrane Ca-ATPase

UR - http://www.scopus.com/inward/record.url?scp=27744545616&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=27744545616&partnerID=8YFLogxK

U2 - 10.1042/BJ20051012

DO - 10.1042/BJ20051012

M3 - Article

C2 - 16080782

AN - SCOPUS:27744545616

VL - 391

SP - 687

EP - 692

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 3

ER -