De novo design and molecular assembly of a transmembrane diporphyrin-binding protein complex.
Journal Article (Journal Article)
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.
Full Text
Duke Authors
Cited Authors
- Korendovych, IV; Senes, A; Kim, YH; Lear, JD; Fry, HC; Therien, MJ; Blasie, JK; Walker, FA; Degrado, WF
Published Date
- November 2010
Published In
Volume / Issue
- 132 / 44
Start / End Page
- 15516 - 15518
PubMed ID
- 20945900
Pubmed Central ID
- PMC3016712
Electronic International Standard Serial Number (EISSN)
- 1520-5126
International Standard Serial Number (ISSN)
- 0002-7863
Digital Object Identifier (DOI)
- 10.1021/ja107487b
Language
- eng