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Lawrence SH , Ramirez UD , Selwood T , Stith L , Jaffe EK
Allosteric inhibition of human porphobilinogen synthase
J Biol Chem. 2009 Dec 18;284(51) :35807-17
AbstractPorphobilinogen synthase (PBGS) catalyzes the first common step in tetrapyrrole (e.g. heme, chlorophyll) biosynthesis. Human PBGS exists as an equilibrium of high activity octamers, low activity hexamers, and alternate dimer configurations that dictate the stoichiometry and architecture of further assembly. It is posited that small molecules can be found that inhibit human PBGS activity by stabilizing the hexamer. Such molecules, if present in the environment, could potentiate disease states associated with reduced PBGS activity, such as lead poisoning and ALAD porphyria, the latter of which is associated with human PBGS variants whose quaternary structure equilibrium is shifted toward the hexamer (Jaffe, E. K., and Stith, L. (2007) Am. J. Hum. Genet. 80, 329-337). Hexamer-stabilizing inhibitors of human PBGS were identified using in silico prescreening (docking) of approximately 111,000 structures to a hexamer-specific surface cavity of a human PBGS crystal structure. Seventy-seven compounds were evaluated in vitro; three provided 90-100% conversion of octamer to hexamer in a native PAGE mobility shift assay. Based on chemical purity, two (ML-3A9 and ML-3H2) were subjected to further evaluation of their effect on the quaternary structure equilibrium and enzymatic activity. Naturally occurring ALAD porphyria-associated human PBGS variants are shown to have an increased susceptibility to inhibition by both ML-3A9 and ML-3H2. ML-3H2 is a structural analog of amebicidal drugs, which have porphyria-like side effects. Data support the hypothesis that human PBGS hexamer stabilization may explain these side effects. The current work identifies allosteric ligands of human PBGS and, thus, identifies human PBGS as a medically relevant allosteric enzyme.
NotesLawrence, Sarah H Ramirez, Ursula D Selwood, Trevor Stith, Linda Jaffe, Eileen K P30 CA006927/CA/NCI NIH HHS/United States R01 ES003654/ES/NIEHS NIH HHS/United States R21 AI063324/AI/NIAID NIH HHS/United States T32 CA009035/CA/NCI NIH HHS/United States Research Support, N.I.H., Extramural United States The Journal of biological chemistry J Biol Chem. 2009 Dec 18;284(51):35807-17. Epub .