Structure determination of the apo-form of human inosine 5'-monophosphate dehydrogenase type II

TL;DRAbstract

Human inosine 5'-monophosphate dehydrogenase (IMPDH) controls a key metabolic step in the regulation of cell differentiation and growth.The catalytic reaction involves the NAD-dependent oxidation of inosine 5'-monophosphate (IMP) to xanthosine 5'-monophosphate (XMP), which is the rate-limiting step in the de novo biosynthesis of guanine nucleotides.There are two identifiable isoforms of IMPDH, type I and type II.Type I is a tissue-specific, constituatively expressed protein while the type II isoform is significantly up-regulated in neoplastic and differentiating cells.As a consequence.IMPDH has been validated as a target in antitumor and irnmunosuppressive drug design.A method was developed to obtain rnilligram quantities of the human type II IMPDH isoform by amplification of the coding region cDNA by PCR, recombining the cDNA into a bacterial nigh-level expression plasmid.and large-scale protein purification to obtain between 25 and 30 mg of protein per liter of bacterial culture.Larg

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