An Escherichia coli expression-based method for heme substitution.
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.
Woodward
Joshua J
JJ
Department of Chemistry, Lawrence Berkeley National Laboratory, University of California, Berkeley, 211 Lewis Hall, Berkeley, California 94720, USA.
Martin
Nathaniel I
NI
Marletta
Michael A
MA
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism*; D006420:Hemeproteins / Q000737:chemistry / Q000378:metabolism*; D011166:Porphyrins / Q000737:chemistry / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013997:Time Factors
D006420:Hemeproteins; D011166:Porphyrins