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Campo DC | Valor | Lengua/Idioma |
---|---|---|
dc.contributor.author | Maestro García-Donas, Beatriz | - |
dc.contributor.author | Sanz, Jesús M. | - |
dc.date.accessioned | 2024-01-31T13:47:11Z | - |
dc.date.available | 2024-01-31T13:47:11Z | - |
dc.date.created | 2005 | - |
dc.identifier.citation | Biochem Journal . 2005 Apr 15;387(Pt 2):479-88 | es_ES |
dc.identifier.issn | 2673-6411 | - |
dc.identifier.uri | https://hdl.handle.net/11000/30900 | - |
dc.description.abstract | Choline-binding modules are present in some virulence factors and many other proteins of Streptococcus pneumoniae (Pneumo coccus). The most extensively studied choline-binding module is C-LytA, the C-terminal moiety of the pneumococcal cell-wall amidase LytA. The three-dimensional structure of C-LytA is built up from six loop-hairpin structures forming a left-handed β solenoid with four choline-binding sites. The affinity of C-LytA for choline and other structural analogues allows its use as an efficient fusion tag for single-step purification of hybrid proteins. In the present study, we characterize the folding and stability of C-LytA by chemical and thermal equilibrium denaturation ex periments. Unfolding experiments using guanidinium chloride at pH 7.0 and 20 ◦ C suggest the existence of two partly folded states (I1 and I2) in the following model: N (native)→I1←→ I2. The N→I1 transition is non-co-operative and irreversible, and is signi ficant even in the absence of a denaturant. In contrast, the I1 ←→ 2transition is co-operative and reversible, with an associated freeenergy change ( G0) of 30.9+− 0.8 kJ · mol−1. The residual structure in the I2 state is unusually stable even in 7.4 M guanidinium chloride. Binding of choline stabilizes the structure of the native state, induces its dimerization and prevents the accumulation of the I1 species ([N]2 [I2]2, G0 =50.1+− 0.8 kJ · mol−1). Fluorescence andCDmeasurements, gel-filtration chromatography and limited proteolysis suggest that I1 differs from N in the local unfolding of the N-terminal β-hairpins, and that I2 has a residual structure in the C-terminal region. Thermal denaturation of C-LytA suggests the accumulation of at least the I1 species. These results might pave the way for an effective improvement of its biotechnological applications by protein engineering. | es_ES |
dc.format | application/pdf | es_ES |
dc.format.extent | 10 | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Biochemical Society | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | * |
dc.subject | affinity tag | es_ES |
dc.subject | choline-binding module | es_ES |
dc.subject | C-LytA | es_ES |
dc.subject | partly folded state | es_ES |
dc.subject | protein folding | es_ES |
dc.subject | β-solenoid | es_ES |
dc.title | Accumulation of partly folded states in the equilibrium unfolding of the pneumococcal choline-binding module C-C-LytA | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.contributor.institute | Institutos de la UMH::Instituto de Bioingeniería | es_ES |
dc.relation.publisherversion | https://doi.org/10.1042/BJ20041194 | es_ES |
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