The primary structure of cytochrome c from the nematode Caenorhabditis elegans

The complete amino acid sequence of cytochrome c from the nematode Caenorhabditis elegans was determined. The native protein displays the same spectral properties in the oxidized and reduced states as horse heart cytochrome c. The apoprotein consists of 110 amino acid residues and differs from human cytochrome c by 44 substitutions, one internal deletion, five N-terminal additions and two C-terminal additions. One of the substitutions is the replacement of an ‘invariant’ phenylalanine residue at position 15 by tyrosine. The N-terminal sequence extension contains a short peptide motif, which is highly homologous with a peptide fragment present at the N-terminus of annelid and insect cytochrome c sequences. From the number of amino acid changes and the evolutionary rate of cytochrome c it would appear that nematodes diverged from a line leading to man about 1.4 billion years ago. When similar data based on the amino acid sequences of the histones H1, H2A, H2B and H3 are taken into account, the average estimate is 1.1 +/- 0.1 billion years.

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The semisynthesis of fragments corresponding to residues 66-104 of horse heart cytochrome c

Biochemical Journal ◽

10.1042/bj1790169 ◽

1979 ◽

Vol 179 (1) ◽

pp. 169-182 ◽

Cited By ~ 10

Author(s):

C J Wallace ◽

R E Offord

Keyword(s):

Biological Activity ◽

Amino Acid ◽

Cytochrome C ◽

Amino Acid Sequences ◽

Side Chain ◽

Horse Heart Cytochrome ◽

Natural Sequence ◽

C Terminus ◽

N Terminus ◽

Horse Heart Cytochrome C

We describe the N epsilon-acetimidylation of horse heart cytochrome c with retention of biological activity, the cleavage of the modified protein by CNBr, the separation of the fragments, and their further side-chain protection. We describe the manipulation of the amino acid sequences of the fragments by stepwise semisynthetic methods. We have prepared fragments corresponding to residues 66-78 and 66-79 of the protein, as well as the [Asp66] analogue of fragment 66-79. We have prepared the natural sequence and the [o-fluoro-Phe82] analogue of the fragment corresponding to residues 81-104 of the protein, and the [N epsilon-trifluoroacetyl-Lys79], the [N epsilon-dinitrophenyl-Lys79] and the [S-acetamidomethyl-Cys79] analogues of fragment 79-104, and the [N epsilon-Cbz-Lys81] analogue of fragment 80-104. We have coupled back the fragments of natural sequence to form a semisynthetic fragment corresponding to residues 66-104 of the protein. Modified fragments were also coupled to give analogues of the 66-104-residue sequence. In every case the homoserine residue representing methionine-80 was removed from the C-terminus of the 66-80-residue fragment and replaced by methionine on the N-terminus of the 81-104 residue fragment during the preparation of the fragments for coupling. The semisynthetic fragments are ready for specific deprotection and further coupling. We have coupled one such fragment to the (1-65)-peptide to produce semisynthetic [Hse65]cytochrome c. The product has satisfactory characteristics on chemical analysis, and on assay of its biological activity.

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Amino-Acid Sequence of Horse Heart Cytochrome C : Peptides released by Digestion with Chymotrypsin

Nature ◽

10.1038/1921121a0 ◽

1961 ◽

Vol 192 (4808) ◽

pp. 1121-1123 ◽

Cited By ~ 109

Author(s):

E. MARGOLIASH ◽

EMIL L. SMITH

Keyword(s):

Amino Acid ◽

Cytochrome C ◽

Amino Acid Sequence ◽

Horse Heart Cytochrome ◽

Horse Heart Cytochrome C

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Enzyme-assisted semisynthesis of polypeptide active esters and their use

Biochemical Journal ◽

10.1042/bj2490083 ◽

1988 ◽

Vol 249 (1) ◽

pp. 83-88 ◽

Cited By ~ 19

Author(s):

K Rose ◽

C Herrero ◽

A E I Proudfoot ◽

R E Offord ◽

C J A Wallace

Keyword(s):

Amino Acid ◽

Cytochrome C ◽

Amino Acid Ester ◽

Amino Group ◽

Horse Heart Cytochrome ◽

C Terminus ◽

Active Ester ◽

Chemical Coupling ◽

Concentrated Solution ◽

Horse Heart Cytochrome C

A method is described for the preparation of polypeptides activated uniquely at the C-terminus. The polypeptide is incubated in a concentrated solution of an amino acid active ester, the latter having its amino group free but adequately protected by protonation. The amino acid ester is coupled via its amino group to the C-terminus of the polypeptide by enzymic catalysis (reverse proteolysis). The resulting polypeptide C-terminal active ester is then isolated and coupled to a suitable amino component (generally a polypeptide) in a subsequent chemical coupling. The method appears to be generally applicable; fragments of horse heart cytochrome c, and porcine insulin, are used as examples. Two new analogues of cytochrome c have been prepared by using this method, with yields of up to 60% in the final coupling. Scope and limitations of the method are discussed.

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The amino acid sequence of sesame (Sesamum indicum L.) and castor (Ricinus communis L.) cytochrome c

Biochemical Journal ◽

10.1042/bj1210439 ◽

1971 ◽

Vol 121 (3) ◽

pp. 439-446 ◽

Cited By ~ 20

Author(s):

E. W. Thompson ◽

M. Richardson ◽

D. Boulter

Keyword(s):

Amino Acid ◽

Cytochrome C ◽

Mung Bean ◽

Ricinus Communis ◽

Sesamum Indicum ◽

Amino Acid Sequences ◽

Amino Acid Residues ◽

Single Chain ◽

Ricinus Communis L ◽

Lysine Residues

The amino acid sequences of sesame (Sesamum indicum L.) and castor (Ricinus communis L.) cytochrome c were determined by using 1.5μmol of protein from each species. Both molecules consist of a single chain of 111 amino acid residues and are homologous with other mitochondrial cytochrome c molecules. Both have an N-acetylated ‘tail’ of eight amino acids and two ∈-N-trimethyl-lysine residues, as also reported for wheat germ (Delange, Glazer & Smith, 1969) and mung-bean cytochrome c (Thompson, Laycock, Ramshaw & Boulter, 1970). Two different preparations of castor cytochrome c differed by one residue. This was glutamic acid for glutamine in position 100. The results for sesame and castor cytochrome c led to a re-examination and subsequent correction to the N-terminal region of the mung-bean cytochrome c sequence, as given by Thompson et al. (1970).

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Structural analyses of the deduced amino acid sequences of a novel type hemecopper terminal oxidase, cytochrome aco3, from alkalophilic Bacillus YN-2000

Canadian Journal of Microbiology ◽

10.1139/w01-114 ◽

2001 ◽

Vol 47 (12) ◽

pp. 1075-1081 ◽

Cited By ~ 1

Author(s):

Kimitoshi Denda ◽

Akira Oshima ◽

Yoshihiro Fukumori

Keyword(s):

Amino Acid ◽

Cytochrome C ◽

Basic Amino Acid ◽

Structural Features ◽

Thermophilic Bacterium ◽

Amino Acid Sequences ◽

Amino Acid Residues ◽

Gene Encoding ◽

Bacterium Bacillus ◽

Alkalophilic Bacillus

Cytochrome aco3 from a facultatively alkalophilic bacterium, Bacillus YN-2000, was found to be alkaline- and heat-tolerant. To better understand the structural features of Bacillus YN-2000 cytochrome aco3, the gene encoding this enzyme was cloned and sequenced. Nucleotide sequence analyses of the region neighboring the acoI (subunit I) gene revealed that the acoII (subunit II) and acoIII (subunit III) genes were concomitantly clustered upstream and downstream of the acoI gene, respectively, forming an operon with transcriptional polarity. The deduced amino acid sequence of subunit I was highly similar to that of cytochrome caa3 from thermophilic bacterium Bacillus PS3 in which the heme a3 could be replaced with heme o. Furthermore, a marked paucity of basic amino acid residues was found in the cytochrome c-binding subunit II, which might be a result of the adaptation to a highly alkaline external milieu.Key words: cytochrome c oxidase, alkalophile, thermostability, heme o, Bacilli.

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The primary structure of histone H2A from the nematode Caenorhabditis elegans

Biochemical Journal ◽

10.1042/bj2430297 ◽

1987 ◽

Vol 243 (1) ◽

pp. 297-300 ◽

Cited By ~ 4

Author(s):

J R Vanfleteren ◽

S M Van Bun ◽

J J Van Beeumen

Keyword(s):

Amino Acid ◽

Caenorhabditis Elegans ◽

Primary Structure ◽

Amino Acid Residues ◽

Histone H2a ◽

Nematode Caenorhabditis Elegans ◽

Calf Thymus ◽

N Terminus ◽

Lysine Residues ◽

Complete Primary Structure

The complete primary structure of histone H2A from the nematode Caenorhabditis elegans was determined. The amino acid chain consists of 126 amino acid residues and has a blocked N-terminus. By comparison with calf thymus histone H2A, the nematode protein shows five deletions, two insertions and 16 substitutions. Most of the changes occur in the N- and C-terminal regions of the molecule, whereas the central part covering the residues 21-120 is quite well conserved. The lysine residues 5, 8 and 10 were found to be partially acetylated.

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