Optimization of the signal-sequence cleavage site for secretion from Bacillus subtilis of a 34-amino acid fragment of human parathyroid hormone

Gene. 1991 Jun 30;102(2):277-82. doi: 10.1016/0378-1119(91)90090-x.

Abstract

We have effected the secretion from Bacillus subtilis of a 34-amino acid (aa) fragment of human parathyroid hormone (PTH,1-34), using a Bacillus amyloliquefaciens neutral protease signal sequence. The secretion efficiency depended on the aa sequence near the signal-sequence cleavage site. We constructed a series of gene fusions encoding different pairs of aa between the signal sequence and PTH,1-34. There was a correlation between those polypeptides which were efficiently secreted and the potential for a beta-turn in the region just beyond the signal-sequence cleavage site. Based on this correlation, we constructed a gene fusion which specified Gly rather than Ala at the C terminus of the signal sequence, thus creating a beta-turn potential at the end of the signal sequence. The change provided a slight increase in secretion efficiency.

MeSH terms

  • Amino Acid Sequence
  • Bacillus subtilis / genetics*
  • Bacillus subtilis / metabolism
  • Base Sequence
  • DNA
  • Humans
  • Molecular Sequence Data
  • Mutation
  • Parathyroid Hormone / genetics*
  • Parathyroid Hormone / metabolism
  • Protein Conformation
  • Protein Processing, Post-Translational
  • Protein Sorting Signals / genetics*
  • Protein Sorting Signals / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Regulatory Sequences, Nucleic Acid*

Substances

  • Parathyroid Hormone
  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • DNA