Photoinduced electron transfer between cytochrome c peroxidase and yeast cytochrome c labeled at Cys 102 with (4-bromomethyl-4'-methylbipyridine)[bis(bipyridine)]ruthenium2+

Geren, Lois; Hahm, Seung; Durham, Bill; Millett, Francis

doi:10.1021/bi00103a009

Cited by 112 publications

(150 citation statements)

References 38 publications

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“…c to CCP compound I in the complex is too fast to measure by using stopped-flow methods. This result was expected on the basis of photo-induced methods used to follow electron transfer in the noncovalent complex (31,33). In addition, the absorbance at 430 nm first increases owing to the formation of Fe(IV)-O and remains constant while ferrocyt.…”

Section: Resultsmentioning

confidence: 63%

See 1 more Smart Citation

Crystal structure and characterization of a cytochrome c peroxidase–cytochrome c site-specific cross-link

Guo

Bhaskar

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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Section: Resultsmentioning

confidence: 63%

“…Indeed, the reaction between CCP compound I and yeast ferrocyt. c is too fast for conventional stopped-flow spectroscopy (31). As an ''inactive'' control, a CCP mutant where the essential Trp-191 has been converted to Gly was used.…”

Section: Resultsmentioning

confidence: 99%

Crystal structure and characterization of a cytochrome c peroxidase–cytochrome c site-specific cross-link

Guo

Bhaskar

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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“…The synthesis of 4-bromomethyl-4Ј-methyl-2,2Ј-bipyridine (Br-dmbpy) and the assembly of the ruthenium complex [Ru(Brdmbpy)(bpy) 2 ](PF 6 ) 2 was performed as described (22). The ruthenium complex was chemoselectively coupled to the deprotected cysteine residues of MOP2 or MOP3 by adding 13 mg (13.4 mol) of [Ru(Br-dmbpy)(bpy) 2 ](PF 6 ) 2 to a degassed solution of 15 mg (1.3 mol) of MOP2 or MOP3 in 500 l of 3:2 0.15 M sodium phosphate, pH 7.5͞acetonitrile and stirring the solution under argon at room temperature for 14 h. The ruthenium-modified peptides were purified by reversed-phase HPLC.…”

Section: Methodsmentioning

confidence: 99%

Modular synthesis of de novo- designed metalloproteins for light-induced electron transfer

Rau

DeJonge

Haehnel

1998

Proc. Natl. Acad. Sci. U.S.A.

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The design and chemical synthesis of two de novo four-helix bundle proteins is described; each protein has two bound cofactors. Their construction from purified peptides is based on the modular assembly of different amphiphilic helices by chemoselective coupling to a cyclic peptide template. In the hydrophobic interior of the antiparallel four-helix bundle these proteins contain a heme in a binding pocket with two ligating histidine residues. A rutheniumtris(bipyridine) complex is covalently bound to different positions at the hydrophilic side of one of the heme-binding helices. Laser-induced electron transfer across the varied distance through this helix has been studied and compared with a pathway analysis. The UV-visible, CD, and mass spectra are consistent with the structure and orientation predetermined by the template.Redox proteins represent the largest group of enzymes. Their cofactors or redox centers are embedded in structural domains with a wide variety of functions, including light-induced charge separation and electron transport. The size of a domain has come within reach of chemical methods by recent developments in peptide synthesis and chemoselective ligation to link unprotected peptides. The de novo design of redox proteins is an attractive approach to investigate the factors involved in electron transfer (ET) and to create proteins with novel functions. Synthetic peptides have been linked sequentially (1) or assembled by templates like transition metals (2), porphyrins (3), carbohydrates (4), and peptides (5). In particular, a cyclic peptide template with orthogonally protected amino acids provides selective ligation and a well-defined relative orientation of the peptide elements (6). Such a branched design of a template-assembled synthetic protein avoids the folding problem encountered in the association of linear peptides (7) and increases the stability of the folded structure (8). The binding of heme groups to bundles of amphiphilic linear helices has been extensively investigated (9, 10). For studies of the ET, different cofactors have been bound to a single helix (11) or bundles of two (12) and three helices (13). Light-induced ET through metalloproteins like cytochromes and myoglobins (14) has been investigated after modification of surface-exposed amino acid residues by ruthenium complexes. Electron tunneling through a protein (15) is a matter of discussion and has been approximated by a homogeneous barrier to ET in complexes like photosynthetic reaction centers (16). However, different regions of a given protein can indeed display different barriers to ET, as shown by an analysis of pathways through the protein (17). We have extended our model of cytochrome b termed MOP1 with a template-based antiparallel four-helix bundle with two different helices (18) to one with three different helices capable of light-induced ET. One of the two heme-binding amphiphilic helices carries besides the ligating histidine a cysteine at the hydrophilic side to which a ruthenium tris-bipyridine comp...

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“…c co-crystal structure 8 together with a wealth of biochemical data indicates that each electron delivered from ferrocyt. c is accepted by the Trp 191 cationic radical 13,14 which explains why Trp191 is essential for activity 15 .…”

Section: Introductionmentioning

confidence: 99%

A Novel Heme and Peroxide-dependent Tryptophan–tyrosine Cross-link in a Mutant of Cytochrome c Peroxidase

Bhaskar

Immoos

Shimizu

et al. 2003

Journal of Molecular Biology

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The crystal structure of a cytochrome c peroxidase mutant where the distal catalytic His52 is converted to a Tyr reveals that the tyrosine side chain forms a covalent bond with the indole ring nitrogen of Trp51. We hypothesize that this novel bond results from peroxide activation by the heme iron followed by oxidation of Trp51 and Tyr52. This hypothesis has been tested by incorporation of a redox-inactive Zn-protoporphyrin into the protein, and the resulting crystal structure shows the absence of a Trp51-Tyr52 crosslink. Instead, the Tyr52 side chain orients away from the heme active site pocket, which requires a substantial rearrangement of residues 72-80 and 134-144. Additional experiments where heme-containing crystals of the mutant were treated with peroxide support our hypothesis that this novel Trp-Tyr cross-link is a peroxide-dependent process mediated by the heme iron. 3 KeywordsCytochrome c peroxidase; X-ray crystal structure; Fe-containing heme; Zn-containing protoporphyrin IX; oxygen radical; tryptophan-tyrosine cross-link; Trp cation radical. 4 AbbreviationsCcP, cytochrome c peroxidase; CcO, cytochrome c oxidase; cyt. c, reduced horse heart or yeast cytochrome c; EDTA, ethylenediaminetetraacetic acid; MPD, 2-methyl-2,4-pentanediol; KPB, potassium phosphate buffer; M r , relative molecular mass; IPTG, isopropylthio--galactoside 5

show abstract

Photoinduced electron transfer between cytochrome c peroxidase and yeast cytochrome c labeled at Cys 102 with (4-bromomethyl-4'-methylbipyridine)[bis(bipyridine)]ruthenium2+

Cited by 112 publications

References 38 publications

Crystal structure and characterization of a cytochrome c peroxidase–cytochrome c site-specific cross-link

Crystal structure and characterization of a cytochrome c peroxidase–cytochrome c site-specific cross-link

Modular synthesis of de novo- designed metalloproteins for light-induced electron transfer

A Novel Heme and Peroxide-dependent Tryptophan–tyrosine Cross-link in a Mutant of Cytochrome c Peroxidase

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