A total solid-phase synthesis of DILP8

Ital J Anat Embryol. 2013;118(1 Suppl):13-4.

Abstract

We have developed a cysteine anchoring method for the synthesis of DILP8 and its analogues. The first is to synthesis of DILP8A SS13-18, C14-MeOBzl, C24-Acm and activate it as DILP8A S13-18, C14-SSPyr C24-Acm. A next step is to synthesize the DILP8BC16-Acm. The desired peptide, DILP8 with Cys(Acm) at A-24 and B-16, was then dissolved in 75% HOAc by addition of Iodine in MeOH and 4M HCl in dioxane. The reaction mixture was monitored by HPLC and the excess iodine was reduced with ascorbic acid. Purification of the peptide was achieved by HPLC. Pure synthetic DILP8 showed a single peak on analytical HPLC with corrected molecular ion. By using the above methods, enough peptide and highly homogenous pure DLP8 were generated.

MeSH terms

  • Animals
  • Cysteine / chemistry
  • Drosophila Proteins / chemical synthesis*
  • Drosophila Proteins / genetics
  • Drosophila Proteins / isolation & purification*
  • Drosophila melanogaster*
  • Humans
  • Intercellular Signaling Peptides and Proteins / chemical synthesis*
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / isolation & purification*
  • Sequence Homology, Amino Acid
  • Solid-Phase Synthesis Techniques / methods*

Substances

  • Drosophila Proteins
  • INSL4 protein, human
  • Intercellular Signaling Peptides and Proteins
  • insulin-like peptide 8, Drosophila
  • Cysteine