Engineering of betabellin-15D: a 64 residue beta sheet protein that forms long narrow multimeric fibrils

Protein Sci. 1998 Jul;7(7):1545-54. doi: 10.1002/pro.5560070708.

Abstract

The betabellin target structure is a beta-sandwich protein consisting of two 32 residue beta-sheets packed against one another by interaction of their hydrophobic faces. The 32 residue chain of betabellin-15S (HSLTAKIpkLTFSIAphTYTCAV pkYTAKVSH, where p=DPro, k=DLys, and h=DHis) did not fold in water at pH 6.5. Air oxidation of betabellin-15S provided betabellin-15D, the 64 residue disulfide bridged two-chain molecule, which also remained unfolded in water at pH 6.5. By circular dichroic spectropolarimetry, the extent of beta structure observed for betabellin-15D increased with the pH and ionic strength of the solution and the betabellin-15D concentration. By electron microscopy, in 5.0 mM MOPS and 0.25 M NaCl at pH 6.9, betabellin-15D formed long narrow multimeric fibrils. A molecular model was constructed to show that the dimensions of these betabellin-15D fibrils are consistent with a single row of beta-sandwich molecules joined by multiple intersheet H-bonds.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Centrifugation
  • Chromatography, High Pressure Liquid
  • Circular Dichroism
  • Copper / metabolism
  • Disulfides
  • Hydrogen Bonding
  • Hydrogen-Ion Concentration
  • Mass Spectrometry
  • Microscopy, Electron
  • Models, Molecular
  • Molecular Sequence Data
  • Osmolar Concentration
  • Peptides
  • Protein Conformation*
  • Protein Denaturation
  • Protein Engineering*
  • Protein Folding*
  • Proteins / chemistry
  • Proteins / isolation & purification
  • Recombinant Proteins

Substances

  • Disulfides
  • Peptides
  • Proteins
  • Recombinant Proteins
  • betabellin 12
  • betabellin 15
  • betabellin 14
  • Copper