BROWSE

Related Scientist

CCS's photo.

CCS
인지 및 사회성 연구단
more info

ITEM VIEW & DOWNLOAD

High-yield synthesis and purification of recombinant human GABA transaminase for high-throughput screening assays

Cited 0 time in webofscience Cited 0 time in scopus
15 Viewed 0 Downloaded
Title
High-yield synthesis and purification of recombinant human GABA transaminase for high-throughput screening assays
Author(s)
Mingu Gordon Park; Ah-reum Han; Su Yeon Kim; Tai Young Kim; 김호민; C. Justin Lee
Publication Date
2021-01
Journal
Journal of Enzyme Inhibition and Medicinal Chemistry, v.36, no.1, pp.2016 - 2024
Publisher
Taylor and Francis Ltd.
Abstract
© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.Many studies have focussed on modulating the activity of γ-aminobutyric acid transaminase (GABA-T), a GABA-catabolizing enzyme, for treating neurological diseases, such as epilepsy and drug addiction. Nevertheless, human GABA-T synthesis and purification have not been established. Thus, biochemical and drug design studies on GABA-T have been performed by using porcine GABA-T mostly and even bacterial GABA-T. Here we report an optimised protocol for overexpression of 6xHis-tagged human GABA-T in human cells followed by a two-step protein purification. Then, we established an optimised human GABA-T (0.5 U/mg) activity assay. Finally, we compared the difference between human and bacterial GABA-T in sensitivity to two irreversible GABA-T inhibitors, gabaculine and vigabatrin. Human GABA-T in homodimeric form showed 70-fold higher sensitivity to vigabatrin than bacterial GABA-T in multimeric form, indicating the importance of using human GABA-T. In summary, our newly developed protocol can be an important first step in developing more effective human GABA-T modulators.
URI
https://pr.ibs.re.kr/handle/8788114/10315
DOI
10.1080/14756366.2021.1975697
ISSN
1475-6366
Appears in Collections:
Center for Cognition and Sociality(인지 및 사회성 연구단) > 1. Journal Papers (저널논문)
Pioneer Research Center for Biomolecular and Cellular Structure(바이오분자 및 세포구조 연구단) > Protein Communication Group(단백질 커뮤니케이션 그룹) > 1. Journal Papers (저널논문)
Files in This Item:
There are no files associated with this item.

qrcode

  • facebook

    twitter

  • Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.
해당 아이템을 이메일로 공유하기 원하시면 인증을 거치시기 바랍니다.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse