Synthesis and conformation of the trimeric coiled-coil segment of laminin

Int J Pept Protein Res. 1992 Jul;40(1):72-9. doi: 10.1111/j.1399-3011.1992.tb00107.x.

Abstract

A disulfide linked 95-mer parallel hetero-trimeric active site segment of laminin was designed and synthesised. The three subunits, A (32-mer), B1 (30-mer) and B2 (33-mer), were prepared by Boc-based solid-phase peptide synthesis involving a two-step trimethylsilyl bromide-thioanisole and HF deprotection procedure. The interlinking of the three subunits was accomplished by the stepwise selective formation of two disulfide bridges using air-oxidation and thallium (III) trifluoroacetate oxidation. The conformations of the synthetic peptides were studied by circular dichroism (CD) spectroscopy, showing that the hetero-dimer, B1-B2, one of the homo-dimers, B1-B1, and the trimer are 30 to 40% in the alpha-helical conformation in aqueous buffer. Variable temperature CD studies demonstrated that the trimer is considerably more stable (melting temperature (Tm) = 61 degrees) than the hetero-dimer, B1-B2 (Tm = 36 degrees).

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Disulfides / chemistry
  • Laminin / chemical synthesis*
  • Laminin / chemistry
  • Macromolecular Substances
  • Molecular Sequence Data
  • Protein Conformation
  • Protein Structure, Secondary

Substances

  • Disulfides
  • Laminin
  • Macromolecular Substances