De novo design and molecular assembly of a transmembrane diporphyrin-binding protein complex

Ivan V. Korendovych; Alessandro Senes; Yong Ho Kim; James D. Lear; H. Christopher Fry; Michael J. Therien; J. Kent Blasie; F. Ann Walker; William F. Degrado

doi:10.1021/ja107487b

De novo design and molecular assembly of a transmembrane diporphyrin-binding protein complex

Ivan V. Korendovych, Alessandro Senes, Yong Ho Kim, James D. Lear, H. Christopher Fry, Michael J. Therien, J. Kent Blasie, F. Ann Walker, William F. Degrado

Research output: Contribution to journal › Article › peer-review

99 Scopus citations

Abstract

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^IIIDPP'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 D₂-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.

Original language	English (US)
Pages (from-to)	15516-15518
Number of pages	3
Journal	Journal of the American Chemical Society
Volume	132
Issue number	44
DOIs	https://doi.org/10.1021/ja107487b
State	Published - Nov 10 2010
Externally published	Yes

ASJC Scopus subject areas

Catalysis
General Chemistry
Biochemistry
Colloid and Surface Chemistry

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10.1021/ja107487b

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abstract = "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 (FeIIIDPP'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 D2-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.",

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AU - Korendovych, Ivan V.

AU - Senes, Alessandro

AU - Kim, Yong Ho

AU - Lear, James D.

AU - Fry, H. Christopher

AU - Therien, Michael J.

AU - Blasie, J. Kent

AU - Walker, F. Ann

AU - Degrado, William F.

PY - 2010/11/10

Y1 - 2010/11/10

N2 - 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 (FeIIIDPP'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 D2-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.

AB - 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 (FeIIIDPP'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 D2-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.

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De novo design and molecular assembly of a transmembrane diporphyrin-binding protein complex

Abstract

ASJC Scopus subject areas

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