PRIMARY STRUCTURE OF HYDROGENASE-I FROM CLOSTRIDIUM-PASTEURIANUM

被引:95
作者
MEYER, J [1 ]
GAGNON, J [1 ]
机构
[1] CEN,DBMS BIOL STRUCT,CNRS,URA 1333,F-38041 GRENOBLE,FRANCE
关键词
D O I
10.1021/bi00104a018
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Peptides obtained by cleavage of Clostridium pasteurianum hydrogenase I have been sequenced. The data allowed design of oligonucleotide probes which were used to clone a 2310-bp Sau3A fragment containing the hydrogenase encoding gene. The latter has been sequenced and was found to translate into a protein composed of 574 amino acids (M(r) = 63 836), including 22 cysteines. C. pasteurianum hydrogenase is homologous to, but longer than, the large subunit of Desulfovibrio vulgaris (Hildenborough) [Fe] hydrogenase. It includes an additional N-terminal domain of ca. 110 amino acids which contains eight cysteine residues and which therefore could accommodate two of its postulated four [4Fe-4S] clusters. C. pasteurianum hydrogenase is most similar in length, cysteine positions, and sequence altogether to the translation product of a putative hydrogenase encoding gene from D. vulgaris (Hildenborough). Comparisons of the available [Fe] hydrogenase sequences show that these enzymes constitute a structurally rather homogeneous family. While they differ in the length of their N-termini and in the number of their [4Fe-4S] clusters, they are highly similar in their C-terminal halves, which are postulated to harbor the hydrogen-activating H cluster. Five conserved cysteine residues occurring in this domain are likely ligands of the H cluster. Possible ligation by other residues, and in particular by methionine, is discussed. The comparisons carried out here show that the H clusters most probably possess a common structural framework in all [Fe] hydrogenases. On the basis of the available data on these proteins and on the current developments in iron-sulfur chemistry, the H clusters possibly contain six to eight iron atoms. The large differences between the DNA compositions of C. pasteurianum (30% G + C) and D. vulgaris (65% G + C) result in significant differences, not only in codon usage for a given amino acid but also in the amino acid compositions of their respective hydrogenases.
引用
收藏
页码:9697 / 9704
页数:8
相关论文
共 53 条
[1]   IRON-SULFUR CLUSTERS OF HYDROGENASE-I AND HYDROGENASE-II OF CLOSTRIDIUM-PASTEURIANUM [J].
ADAMS, MWW ;
ECCLESTON, E ;
HOWARD, JB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (13) :4932-4936
[2]   THE STRUCTURE AND MECHANISM OF IRON-HYDROGENASES [J].
ADAMS, MWW .
BIOCHIMICA ET BIOPHYSICA ACTA, 1990, 1020 (02) :115-145
[3]   CLONING, SEQUENCE DETERMINATION, AND EXPRESSION OF THE GENES ENCODING THE SUBUNITS OF THE NICKEL-CONTAINING 8-HYDROXY-5-DEAZAFLAVIN REDUCING HYDROGENASE FROM METHANOBACTERIUM-THERMOAUTOTROPHICUM DELTA-H [J].
ALEX, LA ;
REEVE, JN ;
ORMEJOHNSON, WH ;
WALSH, CT .
BIOCHEMISTRY, 1990, 29 (31) :7237-7244
[4]   RECENT DEVELOPMENTS IN THE FIELD OF IRON-SULFUR PROTEINS [J].
BEINERT, H .
FASEB JOURNAL, 1990, 4 (08) :2483-2491
[5]   PURIFICATION AND PROPERTIES OF HYDROGENASE FROM CLOSTRIDIUM-PASTEURIANUM W5 [J].
CHEN, JS ;
MORTENSO.LE .
BIOCHIMICA ET BIOPHYSICA ACTA, 1974, 371 (02) :283-298
[6]   STRUCTURAL FEATURES OF MULTIPLE NIFH-LIKE SEQUENCES AND VERY BIASED CODON USAGE IN NITROGENASE GENES OF CLOSTRIDIUM-PASTEURIANUM [J].
CHEN, KCK ;
CHEN, JS ;
JOHNSON, JL .
JOURNAL OF BACTERIOLOGY, 1986, 166 (01) :162-172
[7]   MULTIPLE SEQUENCE ALIGNMENT WITH HIERARCHICAL-CLUSTERING [J].
CORPET, F .
NUCLEIC ACIDS RESEARCH, 1988, 16 (22) :10881-10890
[9]  
CRESTFIELD AM, 1963, J BIOL CHEM, V238, P622
[10]   ENDONUCLEASE-III IS AN IRON SULFUR PROTEIN [J].
CUNNINGHAM, RP ;
ASAHARA, H ;
BANK, JF ;
SCHOLES, CP ;
SALERNO, JC ;
SURERUS, K ;
MUNCK, E ;
MCCRACKEN, J ;
PEISACH, J ;
EMPTAGE, MH .
BIOCHEMISTRY, 1989, 28 (10) :4450-4455