Hole Hopping Across a Protein-Protein Interface

We have investigated photoinduced hole hopping in a Pseudomonas aeruginosa azurin mutant Re126WWCu , where two adjacent tryptophan residues (W124 and W122) are inserted between the Cu center and a Re photosensitizer coordinated to a H126 imidazole (Re = Re (H126)(CO) (dmp) , dmp = 4,7-dimethyl-1,10-...

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Published in:The journal of physical chemistry. B Vol. 123; no. 7; p. 1578
Main Authors: Takematsu, Kana, Pospíšil, Petr, Pižl, Martin, Towrie, Michael, Heyda, Jan, Záliš, Stanislav, Kaiser, Jens T, Winkler, Jay R, Gray, Harry B, Vlček, Antonín
Format: Journal Article
Language:English
Published: United States 21.02.2019
ISSN:1520-5207, 1520-5207
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Summary:We have investigated photoinduced hole hopping in a Pseudomonas aeruginosa azurin mutant Re126WWCu , where two adjacent tryptophan residues (W124 and W122) are inserted between the Cu center and a Re photosensitizer coordinated to a H126 imidazole (Re = Re (H126)(CO) (dmp) , dmp = 4,7-dimethyl-1,10-phenanthroline). Optical excitation of this mutant in aqueous media (≤40 μM) triggers 70 ns electron transport over 23 Å, yielding a long-lived (120 μs) Re (H126)(CO) (dmp )WWCu product. The Re126FWCu mutant (F124, W122) is not redox-active under these conditions. Upon increasing the concentration to 0.2-2 mM, {Re126WWCu } and {Re126FWCu } are formed with the dmp ligand of the Re photooxidant of one molecule in close contact (3.8 Å) with the W122' indole on the neighboring chain. In addition, {Re126WWCu } contains an interfacial tryptophan quadruplex of four indoles (3.3-3.7 Å apart). In both mutants, dimerization opens an intermolecular W122' → //*Re ET channel (// denotes the protein interface, *Re is the optically excited sensitizer). Excited-state relaxation and ET occur together in two steps (time constants of ∼600 ps and ∼8 ns) that lead to a charge-separated state containing a Re(H126)(CO) (dmp )//(W122 )' unit; then (Cu )' is oxidized intramolecularly (60-90 ns) by (W122 )', forming Re (H126)(CO) (dmp )WWCu //(Cu )'. The photocycle is closed by ∼1.6 μs Re (H126)(CO) (dmp ) → //(Cu )' back ET that occurs over 12 Å, in contrast to the 23 Å, 120 μs step in Re126WWCu . Importantly, dimerization makes Re126FWCu photoreactive and, as in the case of {Re126WWCu } , channels the photoproduced "hole" to the molecule that was not initially photoexcited, thereby shortening the lifetime of Re (H126)(CO) (dmp )//Cu . Although two adjacent W124 and W122 indoles dramatically enhance Cu → *Re intramolecular multistep ET, the tryptophan quadruplex in {Re126WWCu } does not accelerate intermolecular electron transport; instead, it acts as a hole storage and crossover unit between inter- and intramolecular ET pathways. Irradiation of {Re126WWCu } or {Re126FWCu } also triggers intermolecular W122' → //*Re ET, and the Re(H126)(CO) (dmp )//(W122 )' charge-separated state decays to the ground state by ∼50 ns Re (H126)(CO) (dmp ) → //(W122 )' intermolecular charge recombination. Our findings shed light on the factors that control interfacial hole/electron hopping in protein complexes and on the role of aromatic amino acids in accelerating long-range electron transport.
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ISSN:1520-5207
1520-5207
DOI:10.1021/acs.jpcb.8b11982