Scanning Transmission Electron Microscopy and Small-Angle Scattering Provide Evidence That Native Escherichia coli ClpP Is a Tetradecamer with an Axial Pore

John Flanagan, Joseph S. Wall, Malcolm S. Capel, Dieter K. Schneider, John Shanklin

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43 Scopus citations


The Escherichia coli ATP-dependent caseinolytic protease (Clp) is composed of two distinct subunits; protease, ClpP, and ATPase, ClpA. Active ClpP has been overexpressed to approximately 50% of soluble protein in E. coli, and purified to homogeneity. Direct mass determination of individual particles using scanning transmission electron microscopy (STEM) yields a mean native molecular mass of 305 ± 9 kDa for the ClpP oligomer, suggesting that it has a tetradecameric structure. Small-angle X-ray scattering (SAXS) curves were determined for ClpP in solution at concentrations of 1 - 10 mg/mL. A combination of STEM and SAXS data was used to derive a model for ClpP, comprising a cylindrical oligomer about 100 Å in diameter and about 75 Å in height with an axial pore about 32-36 Å in diameter. The volume of the pore is estimated to be ~70 000 Å 3 , similar in size to those found in chaperone proteins, and is large enough to accommodate unfolded polypeptide chains, although most globular folded proteins would be excluded.

Original languageEnglish (US)
Pages (from-to)10910-10917
Number of pages8
Issue number34
Publication statusPublished - Jan 1 1995


All Science Journal Classification (ASJC) codes

  • Biochemistry

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