Recombinant production of TEV cleaved human parathyroid hormone

• Published in: Journal of peptide science Vol. 19; no. 8; pp. 504 - 510
• Main Authors: Audu, Christopher O., Cochran, Jared C., Pellegrini, Maria, Mierke, Dale F.
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.08.2013; Wiley Subscription Services, Inc
• Subjects: Amino Acid Sequence; Amino acids; Base Sequence; Chromatography, Affinity; circular dichroism; Endopeptidases - chemistry; Escherichia coli; Humans; LC/MS; Molecular Sequence Data; NMR; parathyroid hormone; Parathyroid Hormone - biosynthesis; Parathyroid Hormone - chemistry; Parathyroid Hormone - isolation & purification; Peptides; Protein Structure, Secondary; Proteolysis; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - isolation & purification; recombinant peptide production; TEV protease; Tobacco etch virus; LC/MS; recombinant peptide production; parathyroid hormone; NMR; circular dichroism; TEV protease
• ISSN: 1075-2617; 1099-1387
• DOI: 10.1002/psc.2528

The parathyroid hormone, PTH, is responsible for calcium and phosphate ion homeostasis in the body. The first 34 amino acids of the peptide maintain the biological activity of the hormone and is currently marketed for calcium imbalance disorders. Although several methods for the production of recombinant PTH(1‐34) have been reported, most involve the use of cleavage conditions that result in a modified peptide or unfavorable side products. Herein, we detail the recombinant production of 15N‐enriched human parathyroid hormone, 15N PTH(1‐34), generated via a plasmid vector that gives reasonable yield, low‐cost protease cleavage (leaving the native N‐terminal serine in its amino form), and purification by affinity and size exclusion chromatography. We characterize the product by multidimensional, heteronuclear NMR, circular dichroism, and LC/MS. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd. An 1H,15N‐HSQC spectrum of uniformly 15N‐labeled parathyroid hormone(1‐34) demonstrating the efficiency of recombinant production with a tobacco etch virus cleavable thioredoxin‐fusion construct.