Midpoint reduction potentials and heme binding stoichiometries of de novo proteins from designed combinatorial libraries

David A. Moffet, Jennifer Foley, Michael H. Hecht

Research output: Contribution to journalArticlepeer-review

42 Scopus citations

Abstract

We previously reported the de novo design of combinatorial libraries of proteins targeted to fold into four-helix bundles. The sequences of these proteins were designed using a binary code strategy in which each position in the linear sequence is designated as either polar or nonpolar, but the exact identity of the amino acid at each position is varied combinatorially. We subsequently reported that approximately half of these binary coded proteins were capable of binding heme. These de novo heme-binding proteins showed CO binding characteristics similar to natural heme proteins, and several were active as peroxidases. Here we analyze the midpoint reduction potentials and heme binding stoichiometries of several of these de novo heme proteins. All the proteins bound heme with a 1:1 stoichiometry. The reduction potentials ranged from -112 to -176 mV. We suggest that this represents an estimate of the default range of potentials for heme proteins that have neither been prejudiced by rational design nor selected by evolution.

Original languageEnglish (US)
Pages (from-to)231-239
Number of pages9
JournalBiophysical Chemistry
Volume105
Issue number2-3
DOIs
StatePublished - Sep 1 2003

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Organic Chemistry

Keywords

  • Binary patterning
  • De novo proteins
  • Heme proteins
  • Protein design
  • Protein reduction potential

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