Apomyoglobin as a molecular recognition surface: expression, reconstitution and crystallization of recombinant porcine myoglobin in Escherichia coli

Recombinant porcine myoglobin has been produced in Escherichiacoli using the cII fusion expression system of Nagai and Th?gersen[Nature, 309, 810–812 (1984)]. After processing and reconstitutionwith haem, the protein is gel-electrophoretically and spectrophotometricallyindistinguishable from native pig myoglobin. Large crystalsof both native and recombinant porcine myoglobin were grownfrom 50 mM sodium phosphate, pH 7.1, 80% ammonium sulphate.The crystals belong to space group C2 (a = 156.9 ?, b = 42.0?, c = 92.2 ?, ß = 127.9?) and diffract to a nominal2.5 ? resolution. We plan to explore apomyoglobin as a bindingsurface in studies combining site-directed mutagenesis and X-rayanalysis. These experiments will be extended by studying thebinding of haem analogues to the mutant apoproteins.