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Mutational analysis of conserved AAA + residues in the archaeal Lon protease from Thermoplasma acidophilum
Author(s) -
Besche Henrike,
Tamura Noriko,
Tamura Tomohiro,
Zwickl Peter
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.08.021
Subject(s) - thermoplasma acidophilum , protease , aaa proteins , proteases , biochemistry , escherichia coli , biology , recombinant dna , mutagenesis , atpase , enzyme , mutation , gene
The Lon protease from the archaeon Thermoplasma acidophilum ( Ta Lon) 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, Ta Lon was shown to be membrane‐bound in its native host and also when expressed in Escherichia coli . Recombinant Ta Lon 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.