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Metal ion accessibility of histidine-modified superfolder green fluorescent protein expressed in Escherichia coli

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Abstract

Green fluorescent protein (GFP) is frequently utilized for metal ion detection and quantification. To improve the metal binding potential of GFP, three residues (N146, F165, and L201) were substituted to histidines. Each variant responded differently upon interaction with metal ions. More than 80% of N146H, having the most accessible surface area, could bind to immobilized metal ions. However, only F165H exhibited significant differences in quenching by soluble metal ions (22% fluorescence decrease) in comparison with the template protein (12%). These findings can be utilized for designing GFP variants for metal binding and sensor applications.

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Acknowledgements

NT is grateful for the Royal Golden Jubilee (RGJ) Ph.D. scholarship under V.P. supervision from the Thailand Research Fund (TRF). This project was also supported by the National Science Research Foundation (VR) and an annual budget grant of Mahidol University (Project No. 02012053-0003).

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Authors and Affiliations

  1. Department of Clinical Microbiology, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand

    Natta Tansila, Chartchalerm Isarankura Na-Ayudhya & Virapong Prachayasittikul

  2. Department of Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, Lund, Sweden

    Kristian Becker & Leif Bülow

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  1. Natta Tansila
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  2. Kristian Becker
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  3. Chartchalerm Isarankura Na-Ayudhya
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  4. Virapong Prachayasittikul
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  5. Leif Bülow
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Correspondence to Leif Bülow.

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Tansila, N., Becker, K., Na-Ayudhya, C.I. et al. Metal ion accessibility of histidine-modified superfolder green fluorescent protein expressed in Escherichia coli . Biotechnol Lett 30, 1391–1396 (2008). https://doi.org/10.1007/s10529-008-9692-7

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  • DOI: https://doi.org/10.1007/s10529-008-9692-7

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