De novo design and molecular assembly of a transmembrane diporphyrin-binding protein complex.
Data(s) |
10/11/2010
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Formato |
15516 - 15518 |
Identificador |
http://www.ncbi.nlm.nih.gov/pubmed/20945900 J Am Chem Soc, 2010, 132 (44), pp. 15516 - 15518 http://hdl.handle.net/10161/4046 1520-5126 |
Idioma(s) |
ENG en_US |
Relação |
J Am Chem Soc 10.1021/ja107487b Journal of the American Chemical Society |
Tipo |
Journal Article |
Cobertura |
United States |
Resumo |
The de novo design of membrane proteins remains difficult despite recent advances in understanding the factors that drive membrane protein folding and association. We have designed a membrane protein PRIME (PoRphyrins In MEmbrane) that positions two non-natural iron diphenylporphyrins (Fe(III)DPP's) sufficiently close to provide a multicentered pathway for transmembrane electron transfer. Computational methods previously used for the design of multiporphyrin water-soluble helical proteins were extended to this membrane target. Four helices were arranged in a D(2)-symmetrical bundle to bind two Fe(II/III) diphenylporphyrins in a bis-His geometry further stabilized by second-shell hydrogen bonds. UV-vis absorbance, CD spectroscopy, analytical ultracentrifugation, redox potentiometry, and EPR demonstrate that PRIME binds the cofactor with high affinity and specificity in the expected geometry. |
Palavras-Chave | #Circular Dichroism #Membrane Proteins #Models, Molecular #Multiprotein Complexes #Porphyrins #Protein Binding #Protein Folding |