title: Homology in structural organization between E. coli ClpAP protease and the eukaryotic 26 S proteasome.
abstract: Energy-dependent protein degradation is carried out by large multimeric protein complexes such as the proteasomes of eukaryotic and archaeal cells and the ATP-dependent proteases of eubacterial cells. Clp protease, a major multicomponent protease of Escherichia coli, consists of a proteolytic component, ClpP, in association with an ATP-hydrolyzing, chaperonin-like component, ClpA. To provide a structural basis for understanding the regulation and mechanism of action of Clp protease, we have used negative staining electron microscopy and image analysis to examine ClpA and ClpP separately, as well as active ClpAP complexes. Digitized images of ClpP and ClpA were analyzed using a novel algorithm designed to detect rotational symmetries. ClpP is composed of two rings of seven subunits superimposed in bipolar fashion along the axis of rotational symmetry. This structure is similar to that formed by the beta subunits of the eukaryotic and archaeal proteasomes. In the presence of MgATP, ClpA forms an oligomer with 6-fold symmetry when viewed en face. Side views of ClpA indicate that the subunits are bilobed with the respective domains forming two stacked rings. ClpAP complexes contain a tetradecamer of ClpP flanked at one or both ends with a hexamer of ClpA, resulting in a symmetry mismatch between the axially aligned molecules. Our findings demonstrate that, despite the lack of sequence similarity between ClpAP and proteasomes, these multimeric proteases nevertheless have a profound similarity in their underlying architecture that may reflect a common mechanism of action.
authors.affiliation: Laboratory of Structural Biology, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: M
authors.initials: M
authors.lastname: Kessel
authors.forename: M R
authors.initials: MR
authors.lastname: Maurizi
authors.forename: B
authors.initials: B
authors.lastname: Kim
authors.forename: E
authors.initials: E
authors.lastname: Kocsis
authors.forename: B L
authors.initials: BL
authors.lastname: Trus
authors.forename: S K
authors.initials: SK
authors.lastname: Singh
authors.forename: A C
authors.initials: AC
authors.lastname: Steven
mesh_terms: D000251:Adenosine Triphosphatases; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000737:chemistry; D049071:Endopeptidase Clp; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D005057:Eukaryotic Cells / Q000201:enzymology*; D007091:Image Processing, Computer-Assisted; D008274:Magnesium / Q000737:chemistry; D015394:Molecular Structure; D009154:Mutation; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000648:ultrastructure; D046988:Proteasome Endopeptidase Complex; D011487:Protein Conformation; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D029968:Escherichia coli Proteins; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; C483568:ClpA protease, E coli; D049071:Endopeptidase Clp; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; D000251:Adenosine Triphosphatases; D008274:Magnesium