Assignment of the heme c resonances in the 360 MHz 1H NMR spectra of cytochrome c

Keller, Regula M.; Wüthrich, Kurt

doi:10.1016/0005-2795(78)90564-0

Cited by 156 publications

(56 citation statements)

References 28 publications

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“…It also bears a strong similarity to the cyanide adducts of Met-80-Ala cyt c (34) and MP-8 (37). Species L, alkaline cyt c, and these cyt c derivatives all have heme methyl shifts ranging from 25 to 10 ppm with an order of 8-CH 3 Ͼ 5-CH 3 Ͼ 1-CH 3 Ͼ 3-CH 3 , as well as an upfield-shifted heme ␦-meso resonance (Table 1) (32)(33)(34)(35)(36)(37). Its similarities with the NMR properties of these characterized paramagnetic proteins indicate that the cyt c unfolding intermediate in urea is ligated by the native His-18 and a second ligand with cylindrical symmetry.…”

Section: Detection Of Non-native Heme Resonances In the Presence Of Umentioning

confidence: 88%

“…In native cyt c, the interaction of the axial ligands (His-18 and Met-80) with the iron is the primary factor that determines the asymmetry of unpaired electron spin density on the heme, resulting in the characteristic chemical shift pattern observed for the four heme methyl groups (29)(30)(31)(32). Replacing Met-80 with other ligands such as a lysine (as in the alkaline form of cyt c, ref.…”

Section: Detection Of Non-native Heme Resonances In the Presence Of Umentioning

confidence: 99%

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NMR investigation of ferricytochrome c unfolding: Detection of an equilibrium unfolding intermediate and residual structure in the denatured state

Russell

Melenkivitz

Bren

2000

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

105

152

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Section: Detection Of Non-native Heme Resonances In the Presence Of Umentioning

confidence: 88%

Section: Detection Of Non-native Heme Resonances In the Presence Of Umentioning

confidence: 99%

NMR investigation of ferricytochrome c unfolding: Detection of an equilibrium unfolding intermediate and residual structure in the denatured state

Russell

Melenkivitz

Bren

2000

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

105

152

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“…Later, the unpaired spin density could be assigned to individual pyrrole rings of haem c (Redfield & Gupta, 1971;Keller & Wuthrich, 1978a).…”

Section: Correlation Between the Co-ordination Geometry Of The Axial mentioning

confidence: 99%

Amino acid sequence, haem-iron co-ordination geometry and functional properties of mitochondrial and bacterial c-type cytochromes

Senn

Wüthrich

1985

Quart. Rev. Biophys.

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Cytochromes are found in all biological oxidation Systems which involve transport of reducing equivalents through organized chains of membrane bound intermediates, regardless of the ultimate oxidant (Keilin, 1966; Bartsch, 1978; Meyer & Kamen, 1982). Thus, cytochromes are present not only in the aerobic mitochondrial and bac-terial respiratory chain, but are also found in much more diversified procariotic Systems, including all varieties of facultative anaerobes (nitrate and nitrite reducers), obligate anaerobes (sulphate reducers and phototrophic sulphur bacteria), facultative photoheterotrophes (phototrophic non-sulphur purple bacteria), and the photoautotrophic cyanobacteria (blue-green algae). Among the different types of cytochromes occurring in the cell, the soluble c-type cytochromes (‘class I’, Meyer & Kamen, 1982) are the most abundant and best characterized group of proteins (Bartsch, 1978; Meyer & Kamen, 1982; Dickerson & Timkovitch, 1975; Lemberg & Barrett, 1973; Salemme, 1977; Ferguson-Miller, Brautigan & Margoliash, 1979). The amino acid sequences of more than 80 mitochrondrial and close to 40 bacterial cytochromes c are known (Meyer & Kamen, 1982; Dickerson & Timkovitch, 1975; Schwartz & Dayhoff, 1976; Dayhoff & Barker, 1978).

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“…Stereospecific assignments of the ␥-methyl protons of valines and ␤-methylene protons were made by analyzing HNHB (23), 15 N-edited NOESY-HSQC (24), and DQF-COSY (25) spectra. All proton signals from the heme moiety were assigned according to the procedure of Keller and Wü thrich (26). The signals of carbons attached to heme protons were assigned with a constant time 13 C-1 H HSQC spectrum (27).…”

Section: Methodsmentioning

confidence: 99%

Selected Mutations in a Mesophilic Cytochrome cConfer the Stability of a Thermophilic Counterpart

Hasegawa

Uchiyama

Tanimoto

et al. 2000

Journal of Biological Chemistry

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Mesophilic cytochrome c 551 of Pseudomonas aeruginosa (PA c 551 ) became as stable as its thermophilic counterpart, Hydrogenobacter thermophilus cytochrome c 552 (HT c 552 ), through only five amino acid substitutions. The five residues, distributed in three spatially separated regions, were selected and mutated with reference to the corresponding residues in HT c 552 through careful structure comparison. Thermodynamic analysis indicated that the stability of the quintuple mutant of PA c 551 could be partly attained through an enthalpic factor. The solution structure of the mutant showed that, as in HT c 552 , there were tighter side chain packings in the mutated regions. Furthermore, the mutant had an increased total accessible surface area, resulting in great negative hydration free energy. Our results provide a novel example of protein stabilization in that limited amino acid substitutions can confer the overall stability of a natural highly thermophilic protein upon a mesophilic molecule.Heat-stable proteins from thermophilic bacteria usually exhibit main chain foldings similar to those of mesophilic counterparts. Mutational studies on mesophilic proteins modeled with respect to thermophilic counterparts have proved that specific side chain interactions in the thermophiles are partially responsible for the higher stability (1-3). Thermodynamic analysis has also indicated that a thermophilic protein can be stabilized through global interaction throughout the molecule (4). It remains enigmatic as to how many amino acid substitutions contribute to the stability of a natural thermophilic protein (5). In some cases, a mesophilic protein only acquires the stability of the thermophilic counterpart after substantial exchanges of a linear sequence; groups of individual mutations are not sufficient (6). Multiple mutations in mesophilic proteins that completely increase the stability to the levels of thermophilic counterparts would provide important information about relationships between local side chain interactions and overall protein stability and demonstrate that the thermophilic character can depend on a limited number of strong noncovalent interactions.Cytochrome c is a powerful tool for characterizing protein stability because structural information on a variety of cytochromes c is available, and heterologous expression systems for holopoteins have been established (7). Cytochrome c 551 (PA c 551 )1 from a mesophile, Pseudomonas aeruginosa, and cytochrome c 552 (HT c 552 ) from a thermophile, Hydrogenobacter thermophilus, are 82-and 80-amino acid proteins, respectively, each with a covalently attached heme. These proteins exhibit 56% sequence identity (8) and almost the same main chain foldings (9), but HT c 552 exhibits much higher stability compared with PA c 551 (10). On a structural comparison between HT c 552 (9) and PA c 551 (11), we identified three distal regions responsible for the higher stability of the former (9). The single mutation Val-78 to Ile (V78I), and two double mutations Phe-7 to Ala/Val-13...

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Assignment of the heme c resonances in the 360 MHz 1H NMR spectra of cytochrome c

Cited by 156 publications

References 28 publications

NMR investigation of ferricytochrome c unfolding: Detection of an equilibrium unfolding intermediate and residual structure in the denatured state

NMR investigation of ferricytochrome c unfolding: Detection of an equilibrium unfolding intermediate and residual structure in the denatured state

Amino acid sequence, haem-iron co-ordination geometry and functional properties of mitochondrial and bacterial c-type cytochromes

Selected Mutations in a Mesophilic Cytochrome cConfer the Stability of a Thermophilic Counterpart

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