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Inhibition of calpain but not caspase activity by spectrin fragments

Cellular & Molecular Biology Letters volume 15, pages 395–405 (2010)Cite this article

Abstract

Calpains and caspases are ubiquitous cysteine proteases that are associated with a variety of cellular pathways. Calpains are involved in processes such as long term potentiation, cell motility and apoptosis, and have been shown to cleave non-erythroid (brain) α- and β-spectrin and erythroid β-spectrin. The cleavage of erythroid α-spectrin by calpain has not been reported. Caspases play an important role in the initiation and execution of apoptosis, and have been shown to cleave non-erythroid but not erythroid spectrin. We have studied the effect of spectrin fragments on calpain and caspase activities. The erythroid and non-erythroid spectrin fragments used were from the N-terminal region of α-spectrin, and C-terminal region of β-spectrin, both consisting of regions involved in spectrin tetramer formation. We observed that the all spectrin fragments exhibited a concentration-dependent inhibitory effect on calpain, but not caspase activity. It is clear that additional studies are warranted to determine the physiological significance of calpain inhibition by spectrin fragments. Our findings suggest that calpain activity is modulated by the presence of spectrin partial domains at the tetramerization site. It is not clear whether the inhibitory effect is substrate specific or is a general effect. Further studies of this inhibitory effect may lead to the identification and development of new therapeutic agents specifically for calpains, but not for caspases. Proteins/peptides with a coiled coil helical conformation should be studied for potential inhibitory effects on calpain activity.

Abbreviations

αI-N1:

first 156 amino acid residues of erythroid α-spectrin

αI-N3:

first 368 amino acid residues of erythroid α-spectrin

αII-N1:

first 147 amino acid residues of non-erythroid α-spectrin

αII-N3:

first 359 amino acid residues of non-erythroid α-spectrin

βI-C1:

residues 1898 to 2083 of erythroid β-spectrin

βII-C1:

residues 1906 to 2093 of non-erythroid β-spectrin

Ac-DEVD-pNA:

N-acetyl-Asp-Glu-Val-Asp-p-nitroaniline, a caspase substrate

BSA:

bovine serum albumin

CHAPS:

3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate

DMSO:

dimethyl sulfoxide

DTT:

dithiothreitol

EDTA:

ethylenediamine tetraacetic acid

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

PBS:

5 mM phosphate buffer at pH 7.4 with 150 mM NaCl

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Author notes

  1. Chloe Antoniou & Qufei Li

    Present address: Department of Biochemistry and Molecular Biology, University of Chicago, 929 East 57th Street, Chicago, IL, 60637, USA

Authors and Affiliations

  1. Department of Chemistry, University of Illinois at Chicago, 845 West Taylor Street, M/C 111, Chicago, IL, 60607, USA

    Ramunas Rolius, Chloe Antoniou, Lidia A. Nazarova, Stephen H. Kim, Garrett Cobb, Pooja Gala, Priyanka Rajaram, Qufei Li & Leslie W.-M. Fung

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  1. Ramunas Rolius
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  2. Chloe Antoniou
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Correspondence to Leslie W.-M. Fung.

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Rolius, R., Antoniou, C., Nazarova, L.A. et al. Inhibition of calpain but not caspase activity by spectrin fragments. Cell Mol Biol Lett 15, 395–405 (2010). https://doi.org/10.2478/s11658-010-0015-3

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  • DOI: https://doi.org/10.2478/s11658-010-0015-3

Key words

  • Spectrin
  • Calpain
  • Caspase
  • Calpain inhibition
  • Cysteine protease

Cellular & Molecular Biology Letters

ISSN: 1689-1392

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