RECOMBINANT PERDEUTERATED PROTEIN AS AN EFFICIENT METHOD FOR MAKING UNAMBIGUOUS HEME PROTON RESONANCE ASSIGNMENTS: CYANIDE-LIGATED GLYCERA DIBRANCHIATA MONOMER METHEMOGLOBIN COMPONENT IV AS AN EXAMPLE

摘要

In order to overcome the difficulties of selectively assigning the heme proton resonances in paramagnetic low-spin heme proteins, a method involving perdeuteration of the globin has been developed. This method allows rapid proton assignments of the heme prosthetic group to be made, however its use is restricted to proteins for which a suitable expression system exists. As an example of this method, the process of making complete heme proton assignments of the cyanide-ligated Glycera dibranchiata monomer hemoglobin Component IV in both the naturally protonated, native protein and the recombinant, perdeuterated protein is presented. There are many potential uses of this method aside from heme-containing proteins.