Secondary Structure and Pd(II) Coordination in S-Layer Proteins from Bacillus sphaericus Studied by Infrared and X-Ray Absorption Spectroscopy
| Autor: | Katrin Pollmann, Sonja Selenska-Pobell, Olesya Savchuk, Karim Fahmy, Johannes Raff, Christoph Hennig, Mohamed L. Merroun |
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| Rok vydání: | 2006 |
| Předmět: |
Spectrophotometry
Infrared Biophysics Supramolecular Assemblies Infrared spectroscopy Bacillus nanocluster Bacillus sphaericus Protein Structure Secondary Nanoclusters Bacterial Proteins Spectroscopy Fourier Transform Infrared Side chain Fourier transform infrared spectroscopy Protein secondary structure X-ray absorption spectroscopy Binding Sites Extended X-ray absorption fine structure biology Chemistry Serine Endopeptidases metal binding Spectrometry X-Ray Emission biology.organism_classification XANES EXAFS Crystallography FTIR Metals Spectrophotometry Palladium |
| Zdroj: | Biophysical Journal 91(2006)3, 996-1007 |
| ISSN: | 0006-3495 |
| Popis: | The S-layer of Bacillus sphaericus strain JG-A12, isolated from a uranium-mining site, exhibits a high metal-binding capacity, indicating that it may provide a protective function by preventing the cellular uptake of heavy metals and radionuclides. This property has allowed the use of this and other S-layers as self-assembling organic templates for the synthesis of nanosized heavy metal cluster arrays. However, little is known about the molecular basis of the metal-protein interactions and their impact on secondary structure. We have studied the secondary structure, protein stability, and Pd((II)) coordination in S-layers from the B. sphaericus strains JG-A12 and NCTC 9602 to elucidate the molecular basis of their biological function and of the metal nanocluster growth. Fourier transform infrared spectroscopy reveals similar secondary structures, containing approximately 35% beta-sheets and little helical structure. pH-induced infrared absorption changes of the side-chain carboxylates evidence a remarkably low pK3 in both strains and a structural stabilization when Pd((II)) is bound. The COO(-)-stretching absorptions reveal a predominant Pd((II)) coordination by chelation/bridging by Asp and Glu residues. This agrees with XANES and EXAFS data revealing oxygens as coordinating atoms to Pd((II)). The additional participation of nitrogen is assigned to side chains rather than to the peptide backbone. The topology of nitrogen- and carboxyl-bearing side chains appears to mediate heavy metal binding to the large number of Asp and Glu in both S-layers at particularly low pH as an adaptation to the environment from which the strain JG-A12 has been isolated. These side chains are thus prime targets for the design of engineered S-layer-based nanoclusters. |
| Databáze: | OpenAIRE |
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