期刊
PROTEIN SCIENCE
卷 20, 期 4, 页码 656-669出版社
WILEY
DOI: 10.1002/pro.594
关键词
cadaverine; pH-sensing; periplasmic domain; ToxR-like protein; X-ray structure
资金
- Deutsche Forschungsgemeinschaft [Exc114/1, JU270/5-3]
- BMBF [05 ES3XBA/5]
- Elite Network of Bavaria
The membrane-integral transcriptional activator CadC comprises sensory and transcriptional regulatory functions within one polypeptide chain. Its C-terminal periplasmic domain, CadC(pd), is responsible for sensing of environmental pH as well as for binding of the feedback inhibitor cadaverine. Here we describe the crystal structure of CadC(pd) (residues 188-512) solved at a resolution of 1.8 angstrom via multiple wavelength anomalous dispersion (MAD) using a ReCl62- derivative. CadC(pd) reveals a novel fold comprising two subdomains: an N-terminal subdomain dominated by a beta-sheet in contact with three alpha-helices and a C-terminal subdomain formed by an eleven-membered alpha-helical bundle, which is oriented almost perpendicular to the helices in the first subdomain. Further to the native protein, crystal structures were also solved for its variants D471N and D471E, which show functionally different behavior in pH sensing. Interestingly, in the heavy metal derivative of CadC(pd) used for MAD phasing a ReCl62- ion was found in a cavity located between the two subdomains. Amino acid side chains that coordinate this complex ion are conserved in CadC homologues from various bacterial species, suggesting a function of the cavity in the binding of cadaverine, which was supported by docking studies. Notably, CadC(pd) forms a homodimer in solution, which can be explained by an extended, albeit rather polar interface between two symmetry-related monomers in the crystal structure. The occurrence of several acidic residues in this region suggests protonation-dependent changes in the mode of dimerization, which could eventually trigger transcriptional activation by CadC in the bacterial cytoplasm.
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