Cryo-EM Structures of Azospirillum brasilense Glutamate Synthase in Its Oligomeric Assemblies

Abstract

Bacterial NADPH-dependent glutamate synthase (GltS) is a complex iron–sulfur flavoprotein that catalyzes the reductive synthesis of two L-Glu molecules from L-Gln and 2-oxo-glutarate. GltS functional unit hosts an α-subunit (αGltS) and a β-subunit (βGltS) that assemble in different αβ oligomers in solution. Here, we present the cryo-electron microscopy structures of Azospirillum brasilense GltS in four different oligomeric states (α4β3, α4β4, α6β4 and α6β6, in the 3.5- to 4.1-Å resolution range). Our study provides a comprehensive GltS model that details the inter-protomeric assemblies and allows unequivocal location of the FAD cofactor and of two electron transfer [4Fe–4S]+1,+2 clusters within βGltS.