Role of μ-calpain in proteolytic cleavage of brain l-glutamic acid decarboxylase

Di Sha, Ying Jin, Heng Wu, Jianning Wei, Chun Hua Lin, Yi Hsuan Lee, Chandana Buddhala, Shafi Kuchay, Athar H. Chishti, Jang Yen Wu

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Glutamic acid decarboxylase (GAD) is the rate-limiting enzyme for γ-aminobutyric acid (GABA) biosynthesis. Previously, we reported the presence of truncated forms of GAD in vivo and in vitro. In addition, an unidentified endogenous protease responsible for proteolytic cleavage of full-length GAD (fGAD) to its truncated form (tGAD) was also observed. In this communication, we report that μ-calpain is a good candidate for conversion of fGAD67 to tGAD67. This conclusion is based on the following observations: 1. purified recombinant GAD67 is cleaved by μ-calpain at specific sites; 2. in brain synaptosomal preparation, GAD67 is cleaved to its truncated form by an endogenous protease which is inhibited by specific calpain inhibitors; 3. in μ-calpain knockout mice, the level of tGAD in the brain is greatly reduced compared with the wild type; 4. when μ-calpain gene is silenced by siRNA, the level of tGAD is also markedly reduced compared to the control group; and 5. μ-calpain is activated by neuronal stimulation and Ca2+-influx. The physiological significance of calpain in regulation of GABA synthesis and GABAergic neurotransmission is also discussed.

Original languageEnglish
Pages (from-to)9-18
Number of pages10
JournalBrain Research
Volume1207
DOIs
Publication statusPublished - May 1 2008

Fingerprint

Glutamate Decarboxylase
Calpain
Brain
gamma-Aminobutyric Acid
Peptide Hydrolases
Aminobutyrates
Knockout Mice
Synaptic Transmission
Small Interfering RNA
Control Groups
Enzymes
Genes

Keywords

  • Calpain
  • GABA synthesis
  • GAD
  • Proteolytic cleavage

ASJC Scopus subject areas

  • Neuroscience(all)
  • Clinical Neurology
  • Developmental Biology
  • Molecular Biology

Cite this

Sha, D., Jin, Y., Wu, H., Wei, J., Lin, C. H., Lee, Y. H., ... Wu, J. Y. (2008). Role of μ-calpain in proteolytic cleavage of brain l-glutamic acid decarboxylase. Brain Research, 1207, 9-18. https://doi.org/10.1016/j.brainres.2008.02.033

Role of μ-calpain in proteolytic cleavage of brain l-glutamic acid decarboxylase. / Sha, Di; Jin, Ying; Wu, Heng; Wei, Jianning; Lin, Chun Hua; Lee, Yi Hsuan; Buddhala, Chandana; Kuchay, Shafi; Chishti, Athar H.; Wu, Jang Yen.

In: Brain Research, Vol. 1207, 01.05.2008, p. 9-18.

Research output: Contribution to journalArticle

Sha, D, Jin, Y, Wu, H, Wei, J, Lin, CH, Lee, YH, Buddhala, C, Kuchay, S, Chishti, AH & Wu, JY 2008, 'Role of μ-calpain in proteolytic cleavage of brain l-glutamic acid decarboxylase', Brain Research, vol. 1207, pp. 9-18. https://doi.org/10.1016/j.brainres.2008.02.033
Sha, Di ; Jin, Ying ; Wu, Heng ; Wei, Jianning ; Lin, Chun Hua ; Lee, Yi Hsuan ; Buddhala, Chandana ; Kuchay, Shafi ; Chishti, Athar H. ; Wu, Jang Yen. / Role of μ-calpain in proteolytic cleavage of brain l-glutamic acid decarboxylase. In: Brain Research. 2008 ; Vol. 1207. pp. 9-18.
@article{98dd3598d39e4597901c728c057ffce4,
title = "Role of μ-calpain in proteolytic cleavage of brain l-glutamic acid decarboxylase",
abstract = "Glutamic acid decarboxylase (GAD) is the rate-limiting enzyme for γ-aminobutyric acid (GABA) biosynthesis. Previously, we reported the presence of truncated forms of GAD in vivo and in vitro. In addition, an unidentified endogenous protease responsible for proteolytic cleavage of full-length GAD (fGAD) to its truncated form (tGAD) was also observed. In this communication, we report that μ-calpain is a good candidate for conversion of fGAD67 to tGAD67. This conclusion is based on the following observations: 1. purified recombinant GAD67 is cleaved by μ-calpain at specific sites; 2. in brain synaptosomal preparation, GAD67 is cleaved to its truncated form by an endogenous protease which is inhibited by specific calpain inhibitors; 3. in μ-calpain knockout mice, the level of tGAD in the brain is greatly reduced compared with the wild type; 4. when μ-calpain gene is silenced by siRNA, the level of tGAD is also markedly reduced compared to the control group; and 5. μ-calpain is activated by neuronal stimulation and Ca2+-influx. The physiological significance of calpain in regulation of GABA synthesis and GABAergic neurotransmission is also discussed.",
keywords = "Calpain, GABA synthesis, GAD, Proteolytic cleavage",
author = "Di Sha and Ying Jin and Heng Wu and Jianning Wei and Lin, {Chun Hua} and Lee, {Yi Hsuan} and Chandana Buddhala and Shafi Kuchay and Chishti, {Athar H.} and Wu, {Jang Yen}",
year = "2008",
month = "5",
day = "1",
doi = "10.1016/j.brainres.2008.02.033",
language = "English",
volume = "1207",
pages = "9--18",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",

}

TY - JOUR

T1 - Role of μ-calpain in proteolytic cleavage of brain l-glutamic acid decarboxylase

AU - Sha, Di

AU - Jin, Ying

AU - Wu, Heng

AU - Wei, Jianning

AU - Lin, Chun Hua

AU - Lee, Yi Hsuan

AU - Buddhala, Chandana

AU - Kuchay, Shafi

AU - Chishti, Athar H.

AU - Wu, Jang Yen

PY - 2008/5/1

Y1 - 2008/5/1

N2 - Glutamic acid decarboxylase (GAD) is the rate-limiting enzyme for γ-aminobutyric acid (GABA) biosynthesis. Previously, we reported the presence of truncated forms of GAD in vivo and in vitro. In addition, an unidentified endogenous protease responsible for proteolytic cleavage of full-length GAD (fGAD) to its truncated form (tGAD) was also observed. In this communication, we report that μ-calpain is a good candidate for conversion of fGAD67 to tGAD67. This conclusion is based on the following observations: 1. purified recombinant GAD67 is cleaved by μ-calpain at specific sites; 2. in brain synaptosomal preparation, GAD67 is cleaved to its truncated form by an endogenous protease which is inhibited by specific calpain inhibitors; 3. in μ-calpain knockout mice, the level of tGAD in the brain is greatly reduced compared with the wild type; 4. when μ-calpain gene is silenced by siRNA, the level of tGAD is also markedly reduced compared to the control group; and 5. μ-calpain is activated by neuronal stimulation and Ca2+-influx. The physiological significance of calpain in regulation of GABA synthesis and GABAergic neurotransmission is also discussed.

AB - Glutamic acid decarboxylase (GAD) is the rate-limiting enzyme for γ-aminobutyric acid (GABA) biosynthesis. Previously, we reported the presence of truncated forms of GAD in vivo and in vitro. In addition, an unidentified endogenous protease responsible for proteolytic cleavage of full-length GAD (fGAD) to its truncated form (tGAD) was also observed. In this communication, we report that μ-calpain is a good candidate for conversion of fGAD67 to tGAD67. This conclusion is based on the following observations: 1. purified recombinant GAD67 is cleaved by μ-calpain at specific sites; 2. in brain synaptosomal preparation, GAD67 is cleaved to its truncated form by an endogenous protease which is inhibited by specific calpain inhibitors; 3. in μ-calpain knockout mice, the level of tGAD in the brain is greatly reduced compared with the wild type; 4. when μ-calpain gene is silenced by siRNA, the level of tGAD is also markedly reduced compared to the control group; and 5. μ-calpain is activated by neuronal stimulation and Ca2+-influx. The physiological significance of calpain in regulation of GABA synthesis and GABAergic neurotransmission is also discussed.

KW - Calpain

KW - GABA synthesis

KW - GAD

KW - Proteolytic cleavage

UR - http://www.scopus.com/inward/record.url?scp=41949097624&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=41949097624&partnerID=8YFLogxK

U2 - 10.1016/j.brainres.2008.02.033

DO - 10.1016/j.brainres.2008.02.033

M3 - Article

C2 - 18377878

AN - SCOPUS:41949097624

VL - 1207

SP - 9

EP - 18

JO - Brain Research

JF - Brain Research

SN - 0006-8993

ER -