Novel inhibitors complexed with glutamate dehydrogenase: allosteric regulation by control of protein dynamics.
Mammalian glutamate dehydrogenase (GDH) is a homohexameric enzyme that catalyzes the reversible oxidative deamination of l-glutamate to 2-oxoglutarate using NAD(P)(+) as coenzyme. Unlike its counterparts from other animal kingdoms, mammalian GDH is regulated by a host of ligands. The recently discovered hyperinsulinism/hyperammonemia disorder showed that the loss of allosteric inhibition of GDH by GTP causes excessive secretion of insulin. Subsequent studies demonstrated that wild-type and hyperinsulinemia/hyperammonemia forms of GDH are inhibited by the green tea polyphenols, epigallocatechin gallate and epicatechin gallate. This was followed by high throughput studies that identified more stable inhibitors, including hexachlorophene, GW5074, and bithionol. Shown here are the structures of GDH complexed with these three compounds. Hexachlorophene forms a ring around the internal cavity in GDH through aromatic stacking interactions between the drug and GDH as well as between the drug molecules themselves. In contrast, GW5074 and bithionol both bind as pairs of stacked compounds at hexameric 2-fold axes between the dimers of subunits. The internal core of GDH contracts when the catalytic cleft closes during enzymatic turnover. None of the drugs cause conformational changes in the contact residues, but all bind to key interfaces involved in this contraction process. Therefore, it seems likely that the drugs inhibit enzymatic turnover by inhibiting this transition. Indeed, this expansion/contraction process may play a major role in the inter-subunit communication and allosteric regulation observed in GDH.
Li
Ming
M
Donald Danforth Plant Science Center, St. Louis, Missouri 63132, USA.
Smith
Christopher J
CJ
Walker
Matthew T
MT
Smith
Thomas J
TJ
D000494:Allosteric Regulation / Q000187:drug effects; D000818:Animals; D001735:Bithionol / Q000737:chemistry / Q000494:pharmacology; D002417:Cattle; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000737:chemistry / Q000494:pharmacology; D006582:Hexachlorophene / Q000737:chemistry / Q000494:pharmacology; D007211:Indoles / Q000737:chemistry* / Q000494:pharmacology; D007700:Kinetics; D009249:NADP / Q000737:chemistry / Q000494:pharmacology; D010636:Phenols / Q000737:chemistry* / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013768:Tetrahymena / Q000201:enzymology
D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D007211:Indoles; D010636:Phenols; D009249:NADP; D006160:Guanosine Triphosphate; D001735:Bithionol; D005969:Glutamate Dehydrogenase; D006582:Hexachlorophene; C489251:5-iodo-3-((3,5-dibromo-4-hydroxyphenyl)methylene)-2-indolinone