An Escherichia coli expression-based method for heme substitution

Nat Methods. 2007 Jan;4(1):43-5. doi: 10.1038/nmeth984. Epub 2006 Dec 24.


Heme reconstitution with porphyrin analogs is a powerful approach toward understanding the molecular function of heme proteins; present methods, however, have not proven to be generally useful. Here we describe the development and application of an expression-based method for introducing modified porphyrins. The approach allows efficient incorporation of heme analogs using a widely available bacterial strain and offers an attractive alternative to present reconstitution methods that subject proteins to harsh, denaturing conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid / methods
  • Escherichia coli / metabolism*
  • Hemeproteins / chemistry
  • Hemeproteins / metabolism*
  • Porphyrins / chemistry
  • Porphyrins / metabolism*
  • Sensitivity and Specificity
  • Spectrophotometry, Ultraviolet / methods
  • Time Factors


  • Hemeproteins
  • Porphyrins