Survival of Helicobacter pylori in acid depends on intrabacterial urease. This urease is a Ni²⁺-containing oligomeric heterodimer. Regulation of its activity and assembly is important for gastric habitation by this neutralophile. The gene complex encodes catalytic subunits (ureA/B), an acid-gated urea channel (ureI) and accessory assembly proteins (ureEFGH). Using yeast-two hybrid analysis (Y2H) for determining protein-protein interactions, UreF as bait identified 4 interacting sequences encoding UreH while, UreG as bait detected, 5 UreE sequences. These results were confirmed by co-immunoprecipitation and ß-galactosidase assays. Native PAGE immunoblotting of H. pylori inner membranes showed interaction of UreA/B with UreI, while UreI deletion mutants lacked this protein interaction. Deletion of ureE, F, G and H did not affect this interaction with UreI. Hence, the accessory proteins UreE/G and UreF/H form dimeric complexes and UreA/B forms a membrane complex with UreI, perhaps enabling assembly of the urease apo-enzyme at the membrane surface and immediate urea access to intra-bacterial urease to allow rapid periplasmic neutralization.