PMID:8977096
| Citation |
Shin, DH, Yoo, SJ, Shim, YK, Seol, JH, Kang, MS and Chung, CH (1996) Mutational analysis of the ATP-binding site in HslU, the ATPase component of HslVU protease in Escherichia coli. FEBS Lett. 398:151-4 |
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| Abstract |
HslU is the ATPase component of the ATP-dependent HslVU protease in Escherichia coli. To gain an insight into the structure and function of HslU, site-directed mutagenesis was performed to generate a mutation in the ATP-binding site of the ATPase (i.e., to replace the Lys63 with Thr). Unlike the wild-type HslU, the mutant form (referred to as HslU/K63T) could not hydrolyze ATP or support the ATP-dependent hydrolysis of N-carbobenzoxy-Gly-Gly-Leu-7-amido-4-methyl coumarin by HslV. The wild-type HslU (a mixture of monomer and dimer) formed a multimer containing 6-8 subunits in the presence of either ATP or ADP, indicating that ATP-binding, but not its hydrolysis, is required for oligomerization of HslU. However, HslU/K63T remained as a monomer whether or not the adenine nucleotides were present. Furthermore, ATP or ADP could protect HslU, but not HslU/K63T, from degradation by trypsin. These results suggest that the mutation in the ATP-binding site results in prevention of the binding of the adenine nucleotides to HslU and hence in impairment of both oligomerization and ATPase function of HslU. |
| Links | |
| Keywords |
ATP-Dependent Proteases; Adenosine Diphosphate/metabolism; Adenosine Triphosphatases/chemistry; Adenosine Triphosphatases/genetics; Adenosine Triphosphatases/metabolism; Adenosine Triphosphate/metabolism; Binding Sites; DNA Mutational Analysis; Endopeptidase Clp; Endopeptidases/chemistry; Endopeptidases/genetics; Endopeptidases/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Heat-Shock Proteins; Mutagenesis, Site-Directed; Oligopeptides/metabolism; Polymers; Serine Endopeptidases/metabolism |
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