Synthesis and characterization of a 25-residue rubredoxin(II)-like metalloprotein and its valine-leucine mutant

FEBS Lett. 1992 Nov 9;312(2-3):219-22. doi: 10.1016/0014-5793(92)80939-e.

Abstract

An iron-sulfur metalloprotein containing the 5-12 and 35-50 residues of Desulfovibrio gigas rubredoxin has been synthesized by Fmoc solid phase peptide synthesis and subsequent peptide folding. A Gly links the two residue chains between Val-5 and Glu-50. Sybyl Tripos structure optimization indicates only minor structural changes of the folded synthetic protein compared to the similar residue positions in the native protein. The UV-VIS spectrum of the reduced synthetic protein is very similar to that of native D. gigas rubredoxin and the molecular mass determined by laser mass spectrometry has the expected value (+/- 2D). No metal is transferred to the gas phase by the laser beam merely by mixing the peptide and iron(II), substantiating that the folding procedure is a necessary pre-requisite for protein formation. The Val-->Leu41 chemical mutant has also been synthesized and behaves in a closely similar fashion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Leucine / genetics
  • Molecular Sequence Data
  • Mutation*
  • Rubredoxins / biosynthesis*
  • Rubredoxins / chemistry
  • Rubredoxins / metabolism
  • Spectrum Analysis
  • Valine / genetics

Substances

  • Rubredoxins
  • Leucine
  • Valine