PMID:15696175
| Citation |
Burton, RE, Baker, TA and Sauer, RT (2005) Nucleotide-dependent substrate recognition by the AAA+ HslUV protease. Nat. Struct. Mol. Biol. 12:245-51 |
|---|---|
| Abstract |
ATP-dependent protein degradation is controlled principally by substrate recognition. The AAA+ HslU ATPase is thought to bind protein substrates, denature them, and translocate the unfolded polypeptide into the HslV peptidase. The lack of well-behaved high-affinity substrates for HslUV (ClpYQ) has hampered understanding of the rules and mechanism of substrate engagement. We show that HslUV efficiently degrades Arc repressor, especially at heat-shock temperatures. Degradation depends on sequences near the N terminus of Arc. Fusion protein and peptide-binding experiments demonstrate that this sequence is a degradation tag that binds directly to HslU. Strong binding of this tag to the enzyme requires ATP and Mg(2+). Furthermore, fusion of this sequence to a protein with marked mechanical stability leads to complete degradation. Thus, these experiments demonstrate that HslUV is a powerful protein unfoldase and that initial substrate engagement by the HslU ATPase must occur after ATP binding. |
| Links |
PubMed Online version:10.1038/nsmb898 |
| Keywords |
Adenosine Triphosphatases/metabolism; Adenosine Triphosphatases/physiology; Adenosine Triphosphate/metabolism; Binding Sites; Endopeptidase Clp/metabolism; Escherichia coli Proteins/metabolism; Heat-Shock Proteins/metabolism; Heat-Shock Proteins/physiology; Magnesium/metabolism; Protein Denaturation/physiology; Protein Folding; Protein Interaction Mapping; Recombinant Fusion Proteins/metabolism; Repressor Proteins/metabolism; Substrate Specificity/physiology; Viral Proteins/metabolism; Viral Regulatory and Accessory Proteins |
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