Expression, purification, and characterization of inosine 5'-monophosphate dehydrogenase from Borrelia burgdorferi.
Inosine 5'-monophosphate dehydrogenase (IMPDH) is the rate-limiting enzyme in de novo guanine nucleotide biosynthesis. IMPDH converts IMP to xanthosine 5'-monophosphate with concomitant conversion of NAD+ to NADH. All IMPDHs characterized to date contain a 130-residue "subdomain" that extends from an N-terminal loop of the alpha/beta barrel domain. The role of this subdomain is unknown. An IMPDH homolog has been cloned from Borrelia burgdorferi, the causative agent of Lyme disease (Margolis, N., Hogan, D., Tilly, K., and Rosa, P. A. (1994) J. Bacteriol. 176, 6427-6432). This homolog has replaced the subdomain with a 50-residue segment of unrelated sequence. We have expressed and characterized the B. burgdorferi IMPDH homolog. This protein has IMPDH activity, which unequivocally demonstrates that the subdomain is not required for catalytic activity. The monovalent cation and dinucleotide binding sites of B. burgdorferi IMPDH are significantly different from those of human IMPDH. Therefore, these sites are targets for the design of specific inhibitors for B. burgdorferi IMPDH. Such inhibitors might be new treatments for Lyme disease.
Zhou
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Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02254, USA.
Cahoon
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M
Rosa
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Hedstrom
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L
D001665:Binding Sites; D015748:Borrelia burgdorferi Group / Q000201:enzymology*; D002414:Cations, Monovalent / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004926:Escherichia coli; D006801:Humans; D007168:IMP Dehydrogenase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008193:Lyme Disease / Q000188:drug therapy; D016133:Polymerase Chain Reaction
D002414:Cations, Monovalent; D004269:DNA, Bacterial; D007168:IMP Dehydrogenase; C037106:endodeoxyribonuclease NdeI; C024388:CTGCAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific