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Molecular cloning, expression, purification, and characterization of shorter forms of human glutamic decarboxylase 67 in an E. coli expression system
Journal article   Open access   Peer reviewed

Molecular cloning, expression, purification, and characterization of shorter forms of human glutamic decarboxylase 67 in an E. coli expression system

Di Sha, Jianning Wei, Heng Wu, Ying Jin and Jang-Yen Wu
Brain research. Molecular brain research, v 136(1)
20 May 2005
DOI: https://doi.org/10.1016/j.molbrainres.2005.02.005
PMID: 15893607
Featured in Collection :   UN Sustainable Development Goals @ Drexel
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https://doi.org/10.1016/j.molbrainres.2005.02.005View
Published, Version of Record (VoR)Maybe Open Access (Publisher Bronze),  Open

Abstract

GABA GABA synthesis GAD l-Glutamic decarboxylase
Previously, we reported the presence of truncated form of human brain l-glutamic decarboxylase 65 (tGAD 65) in vivo as well as in vitro and found that tGAD 65 was more active than the full-length GAD 65 (Wei et al., J. Biomed. Sci., 10: 617–624, 2003). Here, we report the presence of two shorter forms of hGAD 67, namely, hGAD 67 (Δ1–70) and hGAD 67 (Δ1–90), referring to a deletion of 1–70 and 1–90 amino acids from the N-terminal, respectively. The molecular masses of hGAD 67 (Δ1–70) and hGAD 67 (Δ1–90) were found to be 59 kDa and 57 kDa, respectively. Both shorter forms were cloned, expressed, and characterized. In contrast to hGAD 65, the shorter forms of hGAD 67 were much less active than the full-length due to decrease in affinity of PLP towards the shorter enzymes. Both the full-length and one of the shorter forms of GAD 67 were detected in porcine brain extract. Furthermore, the full-length GAD 67 could be converted to both shorter forms by crude brain extract, suggesting that an endogenous protease may be present in the brain, which is responsible for the conversion. The cleavage of GAD 67 seems to be Ca 2+-dependent. The model for the conversion of GAD from full-length GAD to shorter forms of GAD and its physiological implications was proposed.

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