The expression level and cytotoxicity of green fluorescent protein are modulated by an additional N-terminal sequence

Hisao Moriya ^,

Graduate School of Environmental and Life Sciences, Research Core for Interdisciplinary Sciences, Okayama University

Received: 16 February 2020 Accepted: 10 May 2020 Published: 14 May 2020

Abstract
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Nucleotide and amino acid sequences at the N-terminus affect the expression level and cytotoxicity of proteins; however, their effects are not fully understood yet. Here, N-terminal 30 nucleotide/10 amino acid (N10) sequences that affect the expression level and cytotoxicity of a green fluorescent protein were systematically isolated in the budding yeast Saccharomyces cerevisiae. The expression per gene (EPG) and gene copy number limit (CNL) relationships were examined to assess the effects of the N10 sequence. The isolated N10 nucleotide sequences suggested that codon optimality is the major determinant of the protein expression level. A higher number of hydrophobic or cysteine residues in the N10 sequence seemed to increase the cytotoxicity of the protein. Therefore, a high frequency of specific amino acid residues in the outside of the main tertiary structure of proteins might not be preferable.
- green fluorescent protein,
- overexpression,
- expression limit,
- expression level,
- protein cytotoxicity
Citation: Hisao Moriya. The expression level and cytotoxicity of green fluorescent protein are modulated by an additional N-terminal sequence[J]. AIMS Biophysics, 2020, 7(2): 121-132. doi: 10.3934/biophy.2020010

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Abstract

Nucleotide and amino acid sequences at the N-terminus affect the expression level and cytotoxicity of proteins; however, their effects are not fully understood yet. Here, N-terminal 30 nucleotide/10 amino acid (N10) sequences that affect the expression level and cytotoxicity of a green fluorescent protein were systematically isolated in the budding yeast Saccharomyces cerevisiae. The expression per gene (EPG) and gene copy number limit (CNL) relationships were examined to assess the effects of the N10 sequence. The isolated N10 nucleotide sequences suggested that codon optimality is the major determinant of the protein expression level. A higher number of hydrophobic or cysteine residues in the N10 sequence seemed to increase the cytotoxicity of the protein. Therefore, a high frequency of specific amino acid residues in the outside of the main tertiary structure of proteins might not be preferable.

Abbreviation GFP: green fluorescent protein; EPG: expression per gene; CNL: gene copy number limit; SC: synthetic complete;
Acknowledgment

I thank Moriya's lab members for valuable discussions. I especially thank Nozomu Saeki for his help with statistical analyses. This work was partly supported by JSPS KAKENHI grant numbers 18K19300 and 17H03618.

Conflict of interest

The author declares no conflict of interest.

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