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

Published

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

  • 20945900

Electronic International Standard Serial Number (EISSN)

  • 1520-5126

International Standard Serial Number (ISSN)

  • 0002-7863

Digital Object Identifier (DOI)

  • 10.1021/ja107487b

Language

  • eng