The Significance of the Flexible Loop in the Azurin (Az-iso2) from the Obligate Methylotroph Methylomonas sp. Strain J

Inoue, Tsuyoshi; Suzuki, Shinnichiro; Nishio, Nobuya; Yamaguchi, Kazuya; Kataoka, Keisuke; Tobari, Jiro; Xie, Yun; Hamanaka, Sawako; Matsumura, Hideki

doi:10.1016/j.jmb.2003.08.002

Cited by 6 publications

(5 citation statements)

References 45 publications

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“…This can be used to construct cytochrome c variants with targeted properties. Moreover, the results provide an additional evidence of structural and functional significance of the loop domains in protein molecules [57–59]. …”

Section: Resultsmentioning

confidence: 99%

New insight into the mechanism of mitochondrial cytochrome c function

et al. 2017

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We investigate functional role of the P76GTKMIFA83 fragment of the primary structure of cytochrome c. Based on the data obtained by the analysis of informational structure (ANIS), we propose a model of functioning of cytochrome c. According to this model, conformational rearrangements of the P76GTKMIFA83 loop fragment have a significant effect on conformational mobility of the heme. It is suggested that the conformational mobility of cytochrome c heme is responsible for its optimal orientation with respect to electron donor and acceptor within ubiquinol–cytochrome c oxidoreductase (complex III) and cytochrome c oxidase (complex IV), respectively, thus, ensuring electron transfer from complex III to complex IV. To validate the model, we design several mutant variants of horse cytochrome c with multiple substitutions of amino acid residues in the P76GTKMIFA83 sequence that reduce its ability to undergo conformational rearrangements. With this, we study the succinate–cytochrome c reductase and cytochrome c oxidase activities of rat liver mitoplasts in the presence of mutant variants of cytochrome c. The electron transport activity of the mutant variants decreases to different extent. Resonance Raman spectroscopy (RRS) and surface-enhanced Raman spectroscopy (SERS) data demonstrate, that all mutant cytochromes possess heme with the higher degree of ruffling deformation, than that of the wild-type (WT) cytochrome c. The increase in the ruffled deformation of the heme of oxidized cytochromes correlated with the decrease in the electron transport rate of ubiquinol–cytochrome c reductase (complex III). Besides, all mutant cytochromes have lower mobility of the pyrrol rings and methine bridges, than WT cytochrome c. We show that a decrease in electron transport activity in the mutant variants correlates with conformational changes and reduced mobility of heme porphyrin. This points to a significant role of the P76GTKMIFA83 fragment in the electron transport function of cytochrome c.

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Section: Resultsmentioning

confidence: 99%

New insight into the mechanism of mitochondrial cytochrome c function

et al. 2017

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“…The only cupredoxin with a Tyr or Trp residue near the active site is the iso-2 azurin from Methylomonas sp. (strain J) (PDB ID 1CUO); 49 Tyr114 is 5.03 Å from Cu. 50 …”

Section: Are Multicopper Oxidases Protected By Tyr/trp Chains?mentioning

confidence: 99%

Living with Oxygen

2018

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Work on the electronic structures of metal–oxo complexes began in Copenhagen over 50 years ago. This work led to the prediction that tetragonal multiply bonded transition metal–oxos would not be stable beyond the iron–ruthenium–osmium oxo wall in the periodic table and that triply bonded metal–oxos could not be protonated, even in the strongest Brønsted acids. In this theory, only double bonded metal–oxos could attract protons, with basicities being a function of the electron donating ability of ancillary ligands. Such correlations of electronic structure with reactivity have gained importance in recent years, most notably owing to the widespread recognition that high-valent iron–oxos are intermediates in biological reactions critical to life on Earth.In this Account, we focus attention on the oxygenations of inert organic substrates by cytochromes P450, as these reactions involve multiply bonded iron–oxos. We emphasize that P450 iron–oxos are strong oxidants, so strong that they would destroy nearby amino acids if substrates are not oxygenated rapidly; it is our view that these high-valent iron–oxos are such dangerous reactive oxygen species that Nature surely found ways to disable them. Looking more deeply into this matter, mainly by examining many thousands of structures in the Protein Data Bank, we have found that P450s and other enzymes that require oxygen for function have chains of tyrosines and tryptophans that extend from active-site regions to protein surfaces. Tyrosines are near the heme active sites in bacterial P450s, whereas tryptophan is closest in most human enzymes. High-valent iron–oxo survival times taken from hole hopping maps range from a few nanoseconds to milliseconds, depending on the distance of the closest Trp or Tyr residue to the heme. In our proposed mechanism, multistep hole tunneling (hopping) through Tyr/Trp chains guides the damaging oxidizing hole to the protein surface, where it can be quenched by soluble protein or small molecule reductants. As the Earth’s oxygenic atmosphere is believed to have developed about 2.5 billion years ago, the increase in occurrence frequency of tyrosine and tryptophan since the last universal evolutionary ancestor may be in part a consequence of enzyme protective functions that developed to cope with the environmental toxin, O2.

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“…PCC 7120 Pc (2CJ3); (l) Phormidium laminosum Pc (2Q5B); (m) Anabaena variabilis Pc (2GIM); [57] (n) Paracoccus denitrificans amicyanin (Am) (2OV0); (o) Paracoccus versutus Am (1ID2); [58] (p) Paracoccus pantotrophus pseudoazurin (pAz) (3ERX); [59] (q) Alcaligenes faecalis pAz (1PAZ); [60] (r) Methylobacterium extorquens pAz (1PMY); [61] (s) Achromobacter cycloclastes pAz (1BQK); [62] (t) Pseudomonas putida azurin (Az) (1NWO); [63] (u) Pseudomonas fluorescens Az (1JOI); [64] (v) Alcaligenes faecalis Az (2IAA); [65] (w) Pseudomonas aeruginosa Az (5AZU); [66] (x) Achromobacter xylosoxidans Az-II (2CCW); [67] (y) Achromobacter xylosoxidans Az-I (1RKR); [68] (z) Methylomonas sp. J Az (1CUO); [69] (aa) Alcaligenes denitrificans Az (1AZC); [70] (bb) Chloroflexus aurantiacus auracyanin-A (2AAN); (cc) Chloroflexus aurantiacus auracyanin-B (1QHQ); [71] (dd) Acidithiobacillus ferrooxidans rusticyanin (1RCY). [72] …”

Section: Figurementioning

confidence: 99%

The Rise of Radicals in Bioinorganic Chemistry

Gray

2016

Israel Journal of Chemistry

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Prior to 1950, the consensus was that biological transformations occurred in two-electron steps, thereby avoiding the generation of free radicals. Dramatic advances in spectroscopy, biochemistry, and molecular biology have led to the realization that protein-based radicals participate in a vast array of vital biological mechanisms. Redox processes involving high-potential intermediates formed in reactions with O2 are particularly susceptible to radical formation. Clusters of tyrosine (Tyr) and tryptophan (Trp) residues have been found in many O2-reactive enzymes, raising the possibility that they play an antioxidant protective role. In blue copper proteins with plastocyanin-like domains, Tyr/Trp clusters are uncommon in the low-potential single-domain electron-transfer proteins and in the two-domain copper nitrite reductases. The two-domain muticopper oxidases, however, exhibit clusters of Tyr and Trp residues near the trinuclear copper active site where O2 is reduced. These clusters may play a protective role to ensure that reactive oxygen species are not liberated during O2 reduction.

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The Significance of the Flexible Loop in the Azurin (Az-iso2) from the Obligate Methylotroph Methylomonas sp. Strain J

Cited by 6 publications

References 45 publications

New insight into the mechanism of mitochondrial cytochrome c function

New insight into the mechanism of mitochondrial cytochrome c function

Living with Oxygen

The Rise of Radicals in Bioinorganic Chemistry

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