UreE stimulation of GTP-dependent urease activation in the UreD-UreF-UreG-urease apoprotein complex.
The activation of metal-containing enzymes often requires the participation of accessory proteins whose roles are poorly understood. In the case of Klebsiella aerogenes urease, a nickel-containing enzyme, metallocenter assembly requires UreD, UreF, and UreG acting as a protein chaperone complex and UreE serving as a nickel metallochaperone. Urease apoprotein within the UreD-UreF-UreG-urease apoprotein complex is activated to wild-type enzyme activity levels under physiologically relevant conditions (100 microM bicarbonate and 20 microM Ni2+) in a process that requires GTP and UreE. The GTP concentration needed for optimal activation is greatly reduced in the presence of UreE compared to that required in its absence. The amount of UreE provided is critical, with maximal activation observed at a concentration equal to that of Ni2+. On the basis of its ability to facilitate urease activation in the presence of chelators, UreE is proposed to play an active role in transferring Ni2+ to urease apoprotein. Studies involving site-directed variants of UreE provide evidence that His96 has a direct role in metal transfer. The results presented here parallel those obtained from previous in vivo studies, demonstrating the relevance of this in vitro system to the cellular metallocenter assembly process.
Soriano
A
A
Department of Microbiology & Molecular Genetics and Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan 48824, USA.
Colpas
G J
GJ
Hausinger
R P
RP
D001059:Apoproteins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism* / Q000494:pharmacology; D001639:Bicarbonates / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism* / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology; D003300:Copper / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D021902:Enterobacter aerogenes / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D006160:Guanosine Triphosphate / Q000378:metabolism* / Q000494:pharmacology; D007098:Imino Acids / Q000494:pharmacology; D009532:Nickel / Q000494:pharmacology; D009571:Nitrilotriacetic Acid / Q000494:pharmacology; D009876:Operon; D028044:Phosphate-Binding Proteins; D014510:Urease / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015032:Zinc / Q000494:pharmacology
D001059:Apoproteins; D001426:Bacterial Proteins; D001639:Bicarbonates; D002352:Carrier Proteins; D002614:Chelating Agents; D007098:Imino Acids; D028044:Phosphate-Binding Proteins; C086767:UreD protein, Bacteria; C090404:UreE protein, Bacteria; C101639:UreF protein, bacteria; C082270:ureG protein, Bacteria; D003300:Copper; D009532:Nickel; D006160:Guanosine Triphosphate; D014510:Urease; D015032:Zinc; D009571:Nitrilotriacetic Acid; C008109:iminodiacetic acid