Mutational analysis of conserved AAA+ residues in the archaeal Lon protease from Thermoplasma acidophilum.
The Lon protease from the archaeon Thermoplasma acidophilum (TaLon) is composed of an N-terminal ATPase associated with various cellular activities (AAA+) domain and a C-terminal Lon protease domain. Although related in sequence to the soluble Lon proteases, TaLon was shown to be membrane-bound in its native host and also when expressed in Escherichia coli. Recombinant TaLon was purified as a functional high-molecular weight complex displaying ATPase and proteolytic activity. Mutagenesis of conserved AAA+ residues revealed that the Walker A and B motifs, and the sensor 1 and sensor 2' residues were essential for the ATPase activity, while the sensor 2 and the arginine finger were involved in activation of the protease domain.
Besche
Henrike
H
Max-Planck-Institute of Biochemistry, Department of Molecular Structural Biology, Am Klopferspitz 18, 82152 Martinsried, Germany.
Tamura
Noriko
N
Tamura
Tomohiro
T
Zwickl
Peter
P
D049069:ATP-Dependent Proteases; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D049070:Protease La; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics*; D013822:Thermoplasma / Q000201:enzymology*
D017931:DNA Primers; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D049069:ATP-Dependent Proteases; D012697:Serine Endopeptidases; C483567:Lon protein, E coli; D049070:Protease La