Native signal peptide of human ERp57 disulfide isomerase mediates secretion of active native recombinant ERp57 protein in yeast Saccharomyces cerevisiae

•Native signal peptide of human ERp57 is correctly processed in S. cerevisiae.•Native sequence human ERp57 is secreted in yeast S. cerevisiae.•Secreted native recombinant human ERp57 is biologically active. Human ERp57 protein is disulfide isomerase, facilitating proper folding of glycoprotein precu...

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Bibliographic Details
Published in:Protein expression and purification 2013-06, Vol.89 (2), p.131-135
Main Authors: Čiplys, Evaldas, Žitkus, Eimantas, Slibinskas, Rimantas
Format: Article
Language:English
Subjects:
Biologically active
Cloning, Molecular - methods
Gene Expression
Genetic Vectors - genetics
Human ERp57
Humans
Native recombinant
Protein Disulfide-Isomerases - chemistry
Protein Disulfide-Isomerases - genetics
Protein Disulfide-Isomerases - isolation & purification
Protein Disulfide-Isomerases - metabolism
Protein Sorting Signals
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Secretion
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Summary:•Native signal peptide of human ERp57 is correctly processed in S. cerevisiae.•Native sequence human ERp57 is secreted in yeast S. cerevisiae.•Secreted native recombinant human ERp57 is biologically active. Human ERp57 protein is disulfide isomerase, facilitating proper folding of glycoprotein precursors in the concert with ER lectin chaperones calreticulin and calnexin. Growing amount of data also associates ERp57 with many different functions in subcellular locations outside the ER. Analysis of protein functions requires substantial amounts of correctly folded, biologically active protein, and in this study we introduce yeast Saccharomyces cerevisiae as a perfect host for production of human ERp57. Our data suggest that native signal peptide of human ERp57 protein is recognized and correctly processed in the yeast cells, which leads to protein secretion. Secreted recombinant ERp57 protein possesses native amino acid sequence and is biologically active. Moreover, secretion allows simple one-step purification of recombinant ERp57 protein with the yields reaching up to 10mg/L.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2013.03.003