Miniaturized metalloproteins: Application to iron-sulfur proteins

被引:62
作者
Lombardi, A
Marasco, D
Maglio, O
Di Costanzo, L
Nasti, F
Pavone, V
机构
[1] Univ Naples Federico II, Dept Chem, I-80134 Naples, Italy
[2] CNR, Ctr Studio Biocristallog, I-80134 Naples, Italy
关键词
D O I
10.1073/pnas.97.22.11922
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The miniaturization process applied to rubredoxins generated a class of peptide-based metalloprotein models, named METP (miniaturized electron transfer protein). The crystal structure of Desulfovibrio vulgaris rubredoxin was selected as a template for the construction of a tetrahedral (S-gamma-Cys)(4) iron-binding site. Analysis of the structure showed that a sphere of 17 Angstrom in diameter, centered on the metal, circumscribes two unconnected approximately Ct symmetry related beta -hairpins, each containing the -Cys-(Aaa)(2)-Cys-sequence. These observations provided a starting point for the design Of an undecapeptide, which self assembles in the presence of tetrahedrally coordinating metal ions. The METP peptide was synthesized in good yield by standard methodologies. Successful assembly of the METP peptide with Co(II), Zn(II), Fe(II/III), in the expected 2:1 stoichiometry, was proven by UV-visible and circular dichroism spectroscopies. UV-visible analysis of the metal complexes indicated the four Cys ligands tetrahedrally arrange around the metal ion, as designed. Circular dichroism measurements on both the free and metal-bound forms revealed that the metal coordination drives the peptide chain to fold into a turned conformation. NMR characterization of the Zn(II)-METP complex fully supported the structure of the designed model. These results prove that METP reproduces the main features of rubredoxin.
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页码:11922 / 11927
页数:6
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