de Cima…Rubio {Sci Rep 5: 16950}
Tunnel within human carbamoyl phosphate synthetase. This multi-domain enzyme, key for detoxifying ammonia as urea, makes carbamoyl phosphate in three steps (including two phosphorylations), uses two catalytic centres and requires allosteric control by N-acetylglutamate (NAG)
Structure of human carbamoyl phosphate synthetase: deciphering the on/off switch of human ureagenesis
de Cima S, Polo LM, Díez-Fernández C, Martínez AI, Cervera J, Fita I, Rubio V.
Sci Rep 2015 Nov; 5:16950.
Human carbamoyl phosphate synthetase (CPS1), a 1500-residue multidomain enzyme, catalyzes the first step of ammonia detoxification to urea requiring N-acetyl-L-glutamate (NAG) as essential activator to prevent ammonia/amino acids depletion. Here we present the crystal structures of CPS1 in the absence and in the presence of NAG, clarifying the on/off-switching of the urea cycle by NAG. By binding at the C-terminal domain of CPS1, NAG triggers long-range conformational changes affecting the two distant phosphorylation domains. These changes, concerted with the binding of nucleotides, result in a dramatic remodeling that stabilizes the catalytically competent conformation and the building of the ~35 Å-long tunnel that allows migration of the carbamate intermediate from its site of formation to the second phosphorylation site, where carbamoyl phosphate is produced. These structures allow rationalizing the effects of mutations found in patients with CPS1 deficiency (presenting hyperammonemia, mental retardation and even death), as exemplified here for some mutations.
PubMed: 26592762. Doi: 10.1038/srep16950
