Nature Methods
- 4, 43 - 45 (2007)
Published online: 24 December 2006; | doi:10.1038/nmeth984
An Escherichia coli expression–based method for heme substitutionJoshua J Woodward1, Nathaniel I Martin1 & Michael A Marletta1, 2, 31
Department of Chemistry, Lawrence Berkeley National Laboratory, University of California, Berkeley, 211 Lewis Hall, Berkeley, California 94720, USA. 2
Department of Molecular and Cell Biology, Lawrence Berkeley National Laboratory, University of California, Berkeley, 211 Lewis Hall, Berkeley, California 94720, USA. 3
Division of Physical Biosciences, Lawrence Berkeley National Laboratory, University of California, Berkeley, 211 Lewis Hall, Berkeley, California 94720, USA.
Correspondence should be addressed to Michael A Marletta marletta@berkeley.edu 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.
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