Cloning and sequencing of the genes encoding the large and small subunits of the periplasmic (NiFeSe) hydrogenase of Desulfovibrio baculatus

Menon, N; Peck, Harry D.; Gall, Jean Le; Przybyla, Alan

doi:10.1128/jb.169.12.5401-5407.1987

Cited by 94 publications

(63 citation statements)

References 41 publications

(23 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…As the N-terminus of the mature protein was identical to the DNA-predicted sequence, it was proposed that C-terminal processing, whereby the His-Val bond in the large subunit of this hydrogenases is hydrolysed by a specific protease, may be an essential step in the formation of active enzyme. Indications for possible processing of the large subunit had earlier been noticed for the enzyme from D. baculatus [139] and for hydrogenases 1 and 2 from E. coli [134]. Evidence for C-terminal processing has now also been obtained for several other Ni-hydrogenases [79,113,141,183].…”

Section: The "Large' or Nickel-binding Subunitmentioning

confidence: 86%

Nickel hydrogenases: in search of the active site

Albracht

1994

Biochimica et Biophysica Acta (BBA) - Bioenergetics

470

479

View full text Add to dashboard Cite

Section: The "Large' or Nickel-binding Subunitmentioning

confidence: 86%

Nickel hydrogenases: in search of the active site

Albracht

1994

Biochimica et Biophysica Acta (BBA) - Bioenergetics

470

479

View full text Add to dashboard Cite

“…cylindrica is substantially different from any of the five hydrogenases whose DNA sequences have been determined thus far ( D . vulgaris [40], D. baculatus [42], D. gigas [43], Bradyrhizohium japonicum [44] and Rhodobacter capsulatus [45]). There appears to be little similarity amongst the Desulfovibrio enzymes but approximately 80% of the amino acids are conserved between the latter two hydrogenases, both of which are membrane-bound uptake hydrogenases [45].…”

Section: Discussionmentioning

confidence: 99%

Soluble hydrogenase of Anabaena cylindrica

Ewart

Reed

Smith

1990

European Journal of Biochemistry

View full text Add to dashboard Cite

A gene potentially encoding a subunit of the soluble hydrogenase of Anabaena cylindrica was isolated from a genomic library by screening with a set of redundant oligonucleotides, the sequence of which was deduced from the amino acid sequence of the purified hydrogenase subunit that catalyses tritium exchange. The nucleotide sequence of the potential gene was determined from two overlapping DNA fragments spanning 7237 bp of the A . cylindrica genome. The region sequenced contained an open reading frame encoding a protein of 383 amino acids with a predicted molecular mass of 41 108 Da. The NH,-terminal amino acid sequence of the purified enzyme, determined by Edman degradation, corresponds exactly with that deduced from the nucleic acid sequence.No significant amino acid or nucleotide similarity is evident between this gene and the periplasmic hydrogenases from three species of Desuljovibrio (D. vulgaris, D. baculatus and D. gigas), or with the membrane-bound 'uptake' hydrogenases of Bradyrhizobium japonicum and Rhodobacter capsulatus. This suggests that the soluble enzyme from cyanobacteria represents a discrete class of hydrogenase.The gene encoding the second subunit (m = 50 kDa) of the soluble hydrogenase, which is required for the catalysis of hydrogen production from dithionite-reduced methyl viologen [Ewart, G. D.

show abstract

“…These results superficially resemble attempts to express the genes for the periplasmic hydrogenase of sulfate-reducing bacteria in E. coli. The small subunit, predominantly in its immature form, was found in the membrane fraction, and the large subunit was present only in the soluble extract (22,29).…”

Section: Notesmentioning

confidence: 99%

Maturation of membrane-bound hydrogenase of Alcaligenes eutrophus H16

Kortlüke

Friedrich

1992

J Bacteriol

View full text Add to dashboard Cite

. 174:6277-6289, 1992). Other genes located in the adjacent pleiotropic region are also required. In the absence of these genes, MBH is synthesized but is catalytically inactive. Immunological analyses revealed that cells containing active MBH produced the small and large subunits of the enzyme in two distinct conformations each; only one of each, presumably the immature form, occurred in cells devoid of MBH activity. The results suggest that the conversion of the two subunits into the catalytically active membraneassociated heterodimer depends on specific maturation processes mediated by hox genes.The gram-negative, facultative chemolithoautotroph Alcaligenes eutrophus H16 (ATCC 17699) contains two hydrogenases. These enzymes catalyze the oxidation of hydrogen coupled to the reduction of various electron acceptors via an energy-conserving mechanism (reviewed in reference 2).Both enzymes are encoded in the cluster of hox genes on the 450-kb megaplasmid pHG1 (reviewed in reference 9). The membrane-bound hydrogenase (MBH) is a heterodimer composed of a small subunit (SSU; apparent molecular mass, 31 kDa) and a large subunit (LSU; apparent molecular mass, 62 kDa) and contains 10 iron atoms and 1 atom of nickel per mol of enzyme (19). The genes encoding the SSU and the LSU (designated hoxK and hoxG, respectively) belong to a complex locus consisting of 11 open reading frames. The derived amino acid sequences of the two subunits of theA. eutrophus MBH show clear-cut homology (60 to 80% identity) to the respective sequences of [NiFe] hydrogenases from various aerobic hydrogen bacteria. Arrays of cysteines are typical of [NiFe] hydrogenases and probably coordinate metal complexes in the redox-active centers of these enzymes (17).Comparison of the derived amino acid sequence of the SSU of the A. eutrophus MBH with the experimentally determined NH2-terminal sequence indicates the presence of a 43-amino-acid leader peptide (17,19). The experimentally determined NH2-terminal sequence of the LSU is-except for the missing initial methionine-colinear with the sequence deduced from the coding region, ruling out a leader peptide for this protein (17,19).To elucidate the functions of the other genes of the hox cluster in the biosynthesis of MBH, mutants defective for lithoautotrophic growth were generated and subjected to biochemical and genetic analyses. These mutants fell into three major classes: (i) mutants that produce catalytically * Corresponding author.

show abstract

Cloning and sequencing of the genes encoding the large and small subunits of the periplasmic (NiFeSe) hydrogenase of Desulfovibrio baculatus

Cited by 94 publications

References 41 publications

Nickel hydrogenases: in search of the active site

Nickel hydrogenases: in search of the active site

Soluble hydrogenase of Anabaena cylindrica

Maturation of membrane-bound hydrogenase of Alcaligenes eutrophus H16

Contact Info

Product

Resources

About