N-Terminal Amino Acid Affects the Translation Efficiency at Lower Temperatures in a Reconstituted Protein Synthesis System

• Published in: International journal of molecular sciences Vol. 25; no. 10; p. 5264
• Main Authors: Fuse-Murakami, Tomoe, Matsumoto, Rena, Kanamori, Takashi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 12.05.2024
• Subjects: Amino Acid Sequence; Amino Acid Substitution; Amino acids; Amino Acids - metabolism; cell-free protein synthesis; Cell-Free System; Cold Temperature; Dihydrofolate reductase; E coli; early elongation; Efficiency; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Genetic translation; N-terminal sequence; Productivity; Protein Biosynthesis; Protein synthesis; Proteins; PURE system; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Temperature; translation efficiency; Japan; early elongation; N-terminal sequence; cell-free protein synthesis; PURE system; translation efficiency
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25105264

The ( )-based protein synthesis using recombinant elements (PURE) system is a cell-free protein synthesis system reconstituted from purified factors essential for translation. The PURE system is widely used for basic and synthetic biology applications. One of the major challenges associated with the PURE system is that the protein yield of the system varies depending on the protein. Studies have reported that the efficiency of translation is significantly affected by nucleotide and amino acid sequences, especially in the N-terminal region. Here, we investigated the inherent effect of various N-terminal sequences on protein synthesis using the PURE system. We found that a single amino acid substitution in the N-terminal region significantly altered translation efficiency in the PURE system, especially at low temperatures. This result gives us useful suggestions for the expression of the protein of interest in vitro and in vivo.