5G1O

Aspartate transcarbamoylase domain of human CAD in apo form

Summary for 5G1O

• Entry DOI: 10.2210/pdb5g1o/pdb
• Related: 5G1N 5G1P
• Descriptor: CAD protein, 1,2-ETHANEDIOL, GLYCEROL, ... (4 entities in total)
• Functional Keywords: transferase, de novo pyrimidine synthesis, transcarbamoylase, transcarbamylase, cad, carbamoyl phosphate synthetase, dihydroorotase, cooperativity
• Biological source: Homo sapiens (human)
• Cellular location: Cytoplasm : P27708
• Total number of polymer chains: 6
• Total formula weight: 209963.06

Authors

Ruiz-Ramos, A.,Grande-Garcia, A.,Moreno-Morcillo, M.D.,Ramon-Maiques, S. (deposition date: 2016-03-29, release date: 2016-06-22, Last modification date: 2024-01-10)

Primary citation

Ruiz-Ramos, A.,Velazquez-Campoy, A.,Grande-Garcia, A.,Moreno-Morcillo, M.,Ramon-Maiques, S.
Structure and Functional Characterization of Human Aspartate Transcarbamoylase, the Target of the Anti-Tumoral Drug Pala.
Structure, 24:1081-, 2016

PubMed Abstract: CAD, the multienzymatic protein that initiates and controls de novo synthesis of pyrimidines in animals, associates through its aspartate transcarbamoylase (ATCase) domain into particles of 1.5 MDa. Despite numerous structures of prokaryotic ATCases, we lack structural information on the ATCase domain of CAD. Here, we report the structure and functional characterization of human ATCase, confirming the overall similarity with bacterial homologs. Unexpectedly, human ATCase exhibits cooperativity effects that reduce the affinity for the anti-tumoral drug PALA. Combining structural, mutagenic, and biochemical analysis, we identified key elements for the necessary regulation and transmission of conformational changes leading to cooperativity between subunits. Mutation of one of these elements, R2024, was recently found to cause the first non-lethal CAD deficit. We reproduced this mutation in human ATCase and measured its effect, demonstrating that this arginine is part of a molecular switch that regulates the equilibrium between low- and high-affinity states for the ligands.

PubMed: 27265852
DOI: 10.1016/J.STR.2016.05.001

Experimental method

X-RAY DIFFRACTION (2.1 Å)