title: Dual role for the glutamine phosphoribosylpyrophosphate amidotransferase ammonia channel. Interdomain signaling and intermediate channeling.
abstract: Glutamine phosphoribosylpyrophosphate (PRPP) amidotransferase catalyzes the first reaction of de novo purine nucleotide synthesis in two steps at two sites. Glutamine is hydrolyzed to glutamate plus NH(3) at an N-terminal glutaminase site, and NH(3) is transferred through a 20-A hydrophobic channel to a distal PRPP site for synthesis of phosphoribosylamine. Binding of PRPP is required to activate the glutaminase site (termed interdomain signaling) to prevent the wasteful hydrolysis of glutamine in the absence of phosphoribosylamine synthesis. Mutations were constructed to analyze the function of the NH(3) channel. In the wild type enzyme, NH(3) derived from glutamine hydrolysis was transferred to the PRPP site, and little or none was released. Replacement of Leu-415 at the PRPP end of the channel with an alanine resulted in a leaky channel and release of NH(3) to the solvent. Mutations in five amino acids that line the channel and two other residues required for the reorganization of phosphoribosyltransferase domain "flexible loop" that leads to formation of the channel perturbed channel function as well as interdomain signaling. The data emphasize the role of the NH(3) channel in coupling interdomain signaling and NH(3) transfer.
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907, USA.
authors.forename: A K
authors.initials: AK
authors.lastname: Bera
authors.forename: J L
authors.initials: JL
authors.lastname: Smith
authors.forename: H
authors.initials: H
authors.lastname: Zalkin
mesh_terms: D000582:Amidophosphoribosyltransferase / Q000235:genetics / Q000378:metabolism*; D000641:Ammonia / Q000378:metabolism*; D004252:DNA Mutational Analysis; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D011487:Protein Conformation; D012267:Ribosemonophosphates / Q000378:metabolism
chemical_list: D007473:Ion Channels; D012267:Ribosemonophosphates; C008891:phosphoribosylamine; D000641:Ammonia; D000582:Amidophosphoribosyltransferase