PMID:15989953
| Citation |
Hinnerwisch, J, Fenton, WA, Furtak, KJ, Farr, GW and Horwich, AL (2005) Loops in the central channel of ClpA chaperone mediate protein binding, unfolding, and translocation. Cell 121:1029-41 |
|---|---|
| Abstract |
The cylindrical Hsp100 chaperone ClpA mediates ATP-dependent unfolding of substrate proteins bearing "tag" sequences, such as the 11-residue ssrA sequence appended to proteins translationally stalled at ribosomes. Unfolding is coupled to translocation through a central channel into the associating protease, ClpP. To explore the topology and mechanism of ClpA action, we carried out chemical crosslinking and functional studies. Whereas a tag from RepA protein crosslinked proximally to the flexible N domains, the ssrA sequence in GFP-ssrA crosslinked distally in the channel to a segment of the distal ATPase domain (D2). Single substitutions placed in this D2 loop, and also in two apparently cooperating proximal (D1) loops, abolished binding of ssrA substrates and unfolded proteins lacking tags and blocked unfolding of GFP-RepA. Additionally, a substitution adjoining the D2 loop allowed binding of ssrA proteins but impaired their translocation. This loop, as in homologous nucleic-acid translocases, may bind substrates proximally and, coupled with ATP hydrolysis, translocate them distally, exerting mechanical force that mediates unfolding. |
| Links |
PubMed Online version:10.1016/j.cell.2005.04.012 |
| Keywords |
Adenosine Triphosphate/metabolism; Amino Acid Sequence/physiology; Binding Sites/physiology; Endopeptidase Clp/chemistry; Endopeptidase Clp/metabolism; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/metabolism; Hydrolysis; Models, Molecular; Molecular Chaperones/chemistry; Molecular Chaperones/metabolism; Protein Binding/physiology; Protein Folding; Protein Structure, Secondary/physiology; Protein Structure, Tertiary/physiology; Protein Transport/physiology; RNA, Bacterial/chemistry; RNA, Bacterial/metabolism |
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