Preparation of a novel 13C-labelled heme protein

Protohemin chloride 90% enriched with 13C in the terminal carbons of the 2-nd and 4-position vinyl side-chains has been synthesized. The 13C-NMR spectrum of carboxymyoglobin reconstituted with the 13C-enriched hemin revealed two signals of nearly equal intensity at 117.6 and 113.4 ppm downfield of tetramethylsilane. The inequivalence of these vinyl carbons must reflect differences in environment imposed by the protein.