한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제51권1호
  • /
  • Pages.37-42
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  • 2023
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  • 1598-642X(pISSN)
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  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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Heterologous Expression of Human SLC1A5v2 as a Functional Glutamine Transporter in Escherichia coli

  • E Young Kim (Division of EcoScience and Interdisciplinary Program of EcoCreative, Graduate School, Ewha Womans University) ;
  • Ji Won Park (Division of EcoScience and Interdisciplinary Program of EcoCreative, Graduate School, Ewha Womans University) ;
  • Ok Bin Kim (Division of EcoScience and Interdisciplinary Program of EcoCreative, Graduate School, Ewha Womans University)
  • 투고 : 2023.01.26
  • 심사 : 2023.02.15
  • 발행 : 2023.03.28
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초록

Neutral and non-essential amino acid, glutamine (Gln), plays an essential role in supplying nitrogen to all the amino acids and nucleotides in the mammalian body. Gln is also the most important carbon source that provides intermediates for gluconeogenesis and fatty acid synthesis and supplements the tricarboxylic acid cycle in fast-growing cancer cells. Among the known 14 Gln transporter genes, soluted carrier family 1 member 5 (SLC1A5) has been reported to be closely associated with cancer cell growth. Three variants (v1, v2, and v3) have been derived from SLC1A5. Here, we established a heterologous gene expression system for the active form of human SLC1A5 variant-2 (hSLC1A5v2) in Escherichia coli. v2 is the smallest variant that has not yet been studied. Four expression systems were investigated: pBAD, pCold, pET, and pQE. We also addressed the problem of codon usage bias. Although pCold and pET overexpressed hSLC1A5v2 in E. coli, they were functionally inactive. hSLC1A5v2 using the pBAD system was able to catalyze the successful transport of Gln, even if it was not highly expressed. Initial activity of hSLC1A5v2 for [14C] Gln uptake in E. coli reached up to 6.73 μmole·min-1·gDW-1 when the cell was induced with 80 mM L-arabinose. In this study, we demonstrated a heterologous expression system for the human membrane protein, SLC1A5, in E. coli. Our results can be used for the functional comparison of SLC1A5 variants (v1, v2, and v3) in future studies, to facilitae the developement of SLC1A5 inhibitors as effective anticancer drugs.

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과제정보

This study was supported by the Basic Science Research Program of the National Research Foundation of Korea [NRF-2019R1A2C1008066 and NRF-2021R1A6A1A10039823]. We would like to thank Editage (www.editage.co.kr) for English language editing.

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