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Journal of Bacteriology, May 1999, p. 2947-2952, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Carboxy-Terminal Processing of the Large Subunit of [Fe]
Hydrogenase from Desulfovibrio desulfuricans ATCC
7757
E. Claude
Hatchikian,1,*
Valérie
Magro,1
Nicole
Forget,1
Yvain
Nicolet,2 and
Juan C.
Fontecilla-Camps2
Unité de Bioénergétique et
Ingéniérie des Protéines, IBSM, CNRS, 13402 Marseilles Cedex 20,1 and Laboratoire de
Cristallographie et de Cristallogénèse des Protéines,
Institut de Biologie Structurale Jean-Pierre Ebel, CEA-CNRS, 38027 Grenoble Cedex 1,2 France
Received 9 December 1998/Accepted 12 February 1999
 |
ABSTRACT |
hydA and hydB, the genes encoding the large
(46-kDa) and small (13.5-kDa) subunits of the periplasmic [Fe]
hydrogenase from Desulfovibrio desulfuricans ATCC 7757, have been cloned and sequenced. The deduced amino acid sequence of the
genes product showed complete identity to the sequence of the
well-characterized [Fe] hydrogenase from the closely related species
Desulfovibrio vulgaris Hildenborough (G. Voordouw and S. Brenner, Eur. J. Biochem. 148:515-520, 1985). The data show that
in addition to the well-known signal peptide preceding the
NH2 terminus of the mature small subunit, the large subunit
undergoes a carboxy-terminal processing involving the cleavage of a
peptide of 24 residues, in agreement with the recently reported data on
the three-dimensional structure of the enzyme (Y. Nicolet, C. Piras, P. Legrand, E. C. Hatchikian, and J. C. Fontecilla-Camps,
Structure 7:13-23, 1999). We suggest that this C-terminal processing
is involved in the export of the protein to the periplasm.
| |
TEXT |
Two main groups of hydrogenase are
present in sulfate-reducing bacteria of the genus
Desulfovibrio, one containing either nickel or nickel and
selenium atoms in addition to iron-sulfur clusters ([NiFe]
hydrogenases and [NiFeSe] hydrogenases) and one, of higher activity,
containing exclusively iron-sulfur clusters ([Fe] hydrogenases)
(8). The widespread nickel-containing hydrogenases have been
intensively studied (2, 24), and the crystal structure of
the [NiFe] hydrogenase of two Desulfovibrio species was
determined (14, 34, 35). In contrast, [Fe] hydrogenases
form a small family of proteins isolated exclusively from anaerobic
microorganisms (1). Both types of hydrogenase from
Desulfovibrio species are heterodimers, which differ in
their metal center composition, amino acid sequences, mechanistic
properties, sensitivity to inhibitors, and immunological properties
(1, 2, 8, 40). They are usually located in the periplasmic
space, where they play a major role in the energy metabolism of these
microorganisms (19, 21, 32). The mature small subunits of
the two types of hydrogenase are preceded by a complex
NH2-terminal sequence (34 amino acid residues for the
[Fe] hydrogenase and 50 amino acid residues for the [NiFe]
hydrogenase), while the large subunits lack an NH2-terminal signal peptide (39, 40). All potential signal sequences from periplasmic hydrogenases contain a unique strictly conserved element (R-R-X-F-X-K), suggesting that these enzymes are exported to the periplasm via an unusual mechanism of membrane translocation (33, 39). In addition to the NH2-terminal signal peptide
of the small subunit, it was shown that the large subunit of [NiFe]
hydrogenase undergoes a C-terminal processing associated with the
incorporation of nickel in the protein, as reported first for the
Azotobacter vinelandii (9) and Escherichia
coli (27) enzymes.
The periplasmic [Fe] hydrogenase of Desulfovibrio
desulfuricans ATCC 7757, which has previously been characterized,
exhibited a molecular mass of 53.5 kDa and comprises two
different subunits of 42.5 and 11 kDa (13). Electron
paramagnetic resonance studies allowed the identification of two
ferredoxin-type [4Fe-4S]1+ clusters and one
atypical cluster (H cluster) involved in H2 activation and
proposed to be a [6Fe-6S]-type cluster (13). The molecular
properties, N-terminal amino acid sequences of both subunits, electron
paramagnetic resonance spectroscopy, and catalytic properties of the
D. desulfuricans ATCC 7757 periplasmic [Fe] hydrogenase
are highly similar to those of the well-characterized [Fe]
hydrogenase from Desulfovibrio vulgaris Hildenborough
(11, 12, 22, 23, 37).
Although it was not clear from the sequence how [Fe] hydrogenase is
exported to the periplasm, since the large subunit lacks a leader
sequence, it was proposed that a single signal peptide containing the
double-arginine motif operates in the export of both subunits (3,
20, 40). The determination of the three-dimensional structure of
D. desulfuricans [Fe] hydrogenase has shown very recently
(18) that the chain tracing of the large subunit in the
electron density map could not be extended beyond Ala397, which is 24 amino acid residues from the C-terminal Ala 421 as determined by gene
sequencing. In the present study, we determined the nucleotide sequence
of the genes encoding the hydrogenase from D. desulfuricans
ATCC 7757 and showed on the basis of accurate determination of the
molecular mass of its large subunit by mass spectrometry and from its
C-terminal amino acid sequence that it lacks a region of 24 C-terminal
amino acids encoded by the gene for the large subunit.
Nucleotide sequence of the [Fe] hydrogenase genes from D. desulfuricans ATCC 7757.
D. desulfuricans ATCC 7757 was grown at 37°C in a basic lactate-sulfate medium (29).
Two sets of degenerate oligonucleotides based on the amino acid
sequence of the N-terminal regions of the large and small subunits of
D. desulfuricans [Fe] hydrogenase (13) were
synthesized: DOP1 (5' ATH GAR TAY GAR ATG CAY AC 3') and DOP2 (5' CAT
RTA RTC YTT DAT YTG YTT 3'). PCR amplification was performed as
previously reported (15). Agarose gel electrophoresis showed
a unique amplification PCR product of about 1,400 bp. PCR products to
be sequenced were directly cloned in the phagemid MOSblue
system from Amersham (15). DNA fragments were sequenced with
an Applied Biosystems 373A apparatus. Since the partial nucleotide sequences of the [Fe] hydrogenase genes from D. desulfuricans ATCC 7757 and D. vulgaris Hildenborough
are highly similar, we used two oligonucleotides, Fe 1 (5' GGG GGT GAC
AGG ATG GTG CAA 3') and Fe 2 (5' GAT CGT GGA CAG GTG CTG AC 3'),
corresponding to the upstream and downstream nucleotide sequences,
respectively, of the [Fe] hydrogenase genes from D. vulgaris Hildenborough, to amplify the D. desulfuricans
[Fe] hydrogenase genes. PCR was performed directly on chromosomal DNA
by using these two facing oligonucleotides. A unique amplification PCR
product of about 2,000 bp was directly sequenced as described above.
The complete nucleotide sequence coding for the [Fe] hydrogenase from
D. desulfuricans ATCC 7757 has been checked by sequencing of
the PCR product on both strands by using primer Fe 3 (5' ACC TCG TGC
TGC CCC GGC TGG 3') for downstream sequencing and primer Fe 4 (5' CCA
GCC GGG GCA GCA CGA GGT 3') for upstream sequencing (Fig.
1).
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FIG. 1.
Nucleotide sequence of the hydA and
hydB structural genes, encoding the large (46-kDa) and small
(13.5-kDa) subunits of the periplasmic [Fe] hydrogenase of D. desulfuricans ATCC 7757, respectively. The amino acid sequence
deduced from the nucleotide sequence is given in the one-letter amino
acid code. Cysteine residues are underlined, and the locations of the
cleavage sites on the large and small subunits are shown ( ).
Possible sequences serving as the promoter ( 10, 35),
ribosome-binding sites (RBS), and transcription terminator (  )
are indicated. Bold amino acids at the beginning of HydB correspond to
the signal sequence. Bold amino acids at the end of HydA correspond to
the precursor C-terminal sequence. The sequences of the primers Fe1 and
Fe2, drawn from the genomic sequence of the closely related strain
D. vulgaris Hildenborough, may not be completely
representative of the genomic sequence of D. desulfuricans
ATCC 7757. The complete nucleotide sequence has been checked by
sequencing of the PCR product on both strands with primer Fe 3 for
downstream sequence and primer Fe 4 for upstream sequencing.
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Analysis of the nucleotide sequence shows the presence of two
contiguous genes, hydA and hydB (Fig. 1). The
hydA gene, of 1,263 bp, coding for the large subunit is
located between ATG at position 161 and TAG at position 1424. The
hydB gene, of 369 bp, corresponding to the small subunit
starts at ATG position 1438 and terminates at TAG position 1807. The
two genes are very close together, with only 11 bp between them. A
putative promoter region consisting of a 10 sequence (GATATT)
and a 35 sequence (TTTCCG) are located 92 to 133 bp
upstream from the ATG of hydA. A first ribosome binding site
(GGAGG) is located 7 bp upstream from the ATG of
hydA, and a second canonical ribosome binding site
(AGGAGG) is also located 7 bp upstream from ATG of
hydB, overlapping the TAG stop codon of hydA. The
two genes are probably expressed as a single transcriptional unit. An
inverted repeated sequence with a 
G of 13 kcal
mol
1 is situated 200 to 250 bp downstream from
hydB. It might constitute a transcription terminator.
The
hydA nucleotide sequence encodes a polypeptide of 420 amino acids with a calculated molecular mass of 45,820 Da, and the
hydB gene encodes a polypeptide of 122 amino acids with a
calculated
molecular mass of 13,493 Da, excluding the
N-formylmethionine.
The amino acid sequence of the
N-terminal extremity of the small
subunit of
D. desulfuricans [Fe] hydrogenase lacks a peptide of
34 amino acids
encoded by the gene for the small subunit (
13),
indicating
that the N-terminal alanine residue of the mature small
subunit
(calculated molecular mass, 10,135 Da) is preceded by
a potential
signal sequence. This N-terminal signal peptide contains
the conserved
feature (RRXFXK) reported for other
Desulfovibrio species
[Fe] hydrogenases (
40).
When the nucleotide sequence of the genes encoding the [Fe]
hydrogenase from
D. desulfuricans ATCC 7757 was compared
with
that of the well-characterized hydrogenase from
D. vulgaris Hildenborough
(
37), only 14 of 1,649 nucleotides were found to be different,
indicating that the two strains
are closely related, in agreement
with the data based on the analysis
of 16S rRNA (
6). Furthermore,
these differences always
affect the third letter of the codons,
leading to total identity
between both pairs of amino acid sequences
(
37) (see below).
As reported for
D. vulgaris Hildenborough
[Fe]
hydrogenase, the three iron-sulfur clusters known to be present
in
D. desulfuricans ATCC 7757 [Fe] hydrogenase
(
13) must all
coordinate to large-subunit cysteine residues,
since the mature
small subunit lacks
cysteine.
Mass spectrometric data.
[Fe] hydrogenase from D. desulfuricans ATCC 7757 was purified and S-carboxymethylated
hydrogenase was prepared as described previously (13). In
this study, matrix-assisted laser desorption/ionization time-of-flight
(MALDI-TOF) mass spectrometry was used to measure the masses of the
subunits of [Fe] hydrogenase with samples of native and
S-carboxymethylated protein. The sample of native [Fe] hydrogenase
(21 µM) was obtained after careful dialysis against distilled water.
Lyophilized S-carboxymethylated [Fe] hydrogenase (4 nmol) was
dissolved in 80 µl of acetic acid and diluted with 200 µl of
distilled water to a solution concentration of 14 µM. A 20-mg/ml
sinapinic acid solution in H2O-0.1%
trifluoroacetate/acetonitrile (60:40) was freshly prepared prior to
experimentation. Then 1.4 µl of a 1:1 mixture of either native [Fe]
hydrogenase or S-carboxymethylated hydrogenase solution and sinapinic
acid solution was applied to the sample plate and the droplets were
allowed to dry at room temperature before insertion. External mass
calibration was provided by the [M+H]+ and
[M+2H]2+ ions of apo-myoglobin (16,951.5 and
8,476.3 Da, respectively). Mass spectrometry was performed with a
Voyager DE-RP (Perspective Biosystems, Framingham, Mass.) MALDI-TOF
mass spectrometer equipped with an XY multisample probe. Ionization was
accomplished with a 337-nm beam from a nitrogen laser with 3-ns-wide
pulses. All data were acquired at 25 kV of acceleration potential in
the positive-ion mode with the linear detector.
The measured mass from the singly charged ion of the
S-carboxymethylated small subunit was found to be 10,133.4 Da (Fig.
2),
in excellent agreement with the value
of 10,135 Da expected from
the predicted amino acid sequence of the
mature small subunit.
The mass spectrum also displays the doubly
charged ion of the
small subunit, corresponding to a mass of 5,068.3 Da. When measurements
were made on native [Fe] hydrogenase (data not
shown), the value
for the small subunit (10,133.7 Da) was found to be
identical
to that obtained with the small subunit of the
S-carboxymethylated
hydrogenase, in agreement with the lack of cysteine
residues and
iron-sulfur clusters in this subunit. The measured mass
for the
singly charged ion of the S-carboxymethylated large subunit of
D. desulfuricans [Fe] hydrogenase was found to be 44,194 Da (Fig.
2). The MALDI-TOF spectrum showed two other charged species
related
to [M+H]
2+ and [2M+H]
+ ions, with
corresponding masses of 22,107 and 88,558 Da, respectively.
The mass of
the large-subunit apoprotein of [Fe] hydrogenase can
be deduced from
the mass of the S-carboxymethylated large subunit,
taking account of
the presence of 18 alkylated cysteine residues
within the protein. A
mass of 43,149 Da is then obtained for the
large subunit by subtracting
the mass of 18 -CH
2-COOH alkyl groups,
each with a
calculated mass of 59 Da, and adding the mass of 18
H atoms.
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FIG. 2.
MALDI-TOF mass spectrum of the S-carboxymethylated
[Fe] hydrogenase of D. desulfuricans. The measured
molecular masses of the S-carboxymethylated small and large subunits
are 10,133.4 and 44,194 Da, respectively.
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|
MALDI-TOF mass spectrometry measurements made on native [Fe]
hydrogenase yielded a mass of 43,316 Da for the large subunit
(data not
shown), a value which is slightly higher than that obtained
for the
large-subunit apoprotein (43,149 Da). This higher molecular
mass could
arise from intermediate protein species which appear
during
decomposition of the iron-sulfur clusters bound to the
native large
subunit. Mass spectrometric analyses on [Fe] hydrogenase
from
D. desulfuricans ATCC 7757 indicated that the measured
molecular
mass of the large-subunit apoprotein (43,149 Da) was smaller
than
the expected molecular mass based on the gene-deduced amino acid
sequence (45,820 Da). The difference (2,671 Da) suggests the occurrence
of carboxy-terminal processing of a precursor form of the large
subunit, since previous studies have shown that the native large
subunit lacks an N-terminal signal sequence (
13).
C-terminal sequence of the large subunit of D. desulfuricans ATCC 7757 [Fe] hydrogenase.
The C-terminal
sequence of the large subunit of the S-carboxymethylated [Fe]
hydrogenase of D. desulfuricans ATCC 7757 was deduced by
digestion with carboxypeptidase Y. A time course analysis of
carboxypeptidase Y digestion revealed only alanine after 15 min. With
increasing times of incubation, other residues, including glutamic
acid, leucine, valine, and glycine, were released in addition to
alanine. The ratio of amino acids released after a 2-h digestion was
Ala (0.96), Glu (0.33), Leu (0.20), Val (0.16), and Gly (0.10). These
data are consistent with the following C-terminal sequence
-Gly-Val-Leu-Glu-Ala.
This amino acid sequence differs from the C-terminal sequence encoded
by the gene for the large subunit (-Ser-Ala-Asn-Lys-Ala421)
(Fig.
1).
The data demonstrated that the carboxyl end of the mature
large subunit
was 24 amino acids shorter than the amino acid sequence
deduced from
the gene and that the C-terminal cleavage occurred
after Ala397 (Fig.
3). It follows that the large subunit of
[Fe]
hydrogenase from
D. desulfuricans ATCC 7757 is
synthesized as
a larger precursor protein from which the mature large
subunit
is derived by proteolytic cleavage of a C-terminal peptide.
Hence,
the mature large subunit is 2,668 Da smaller than the precursor
form of the protein (45,820 Da) calculated from the gene-deduced
amino
acid sequence. This gives a value of 43,152 Da, in good
agreement with
the molecular mass measured by mass spectrometry
for the mature large
subunit (43,149 Da).
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FIG. 3.
Alignment of the gene-deduced C-terminal sequence of the
large subunit of the dimeric [Fe] hydrogenases from
Desulfovibrio species. Dd, [Fe] hydrogenase from D. desulfuricans ATCC 7757 (this work); hydrophobic residues (alanine
and leucine) are in boldface type, and charges are as indicated. DvH,
[Fe] hydrogenase from D. vulgaris Hildenborough
(37). DvoM, [Fe] hydrogenase from D. vulgaris
subsp. oxamicus Monticello (38). Df, [Fe]
hydrogenase from D. fructosovorans (5). The
position of the putative processing site is indicated ( ).
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The precursor C-terminal sequence of the
D. desulfuricans
[Fe] hydrogenase large subunit is highly basic and exhibits a high
percentage of hydrophobic (alanine and leucine) and hydroxylated
amino
acids (Fig.
3). These properties are shared by mitochondrial
targeting
NH
2-terminal sequences, which display, in contrast to
the
precursor C-terminal sequence, a regular spacing of the positive
charges, allowing the formation of an amphiphilic helix (
26,
36). Based on these data and a comparative analysis of the gene
sequences of the large subunits of other dimeric [Fe] hydrogenases
from
Desulfovibrio species which exhibit a high degree of
similarity
(Fig.
3), it can be postulated that the large subunits of
these
enzymes undergo C-terminal processing involving cleavage at a
site analogous to Ala397 of
D. desulfuricans ATCC 7757 hydrogenase
(Fig.
3). As with the NH
2-terminal signal
peptides of small subunits
of [Fe] and [NiFe] hydrogenases
(
40) and cytochrome
c3
(
15),
the peptidase cleavage site would occur after an
alanine residue,
except for the putative C-terminal peptide of
Desulfovibrio fructosovorans [Fe] hydrogenase, which
exhibits a cleavage site located after
a serine residue (
5).
The significance of the C-terminal processing of the large subunit of
[Fe] hydrogenase of
Desulfovibrio species remains to
be
elucidated. It was demonstrated that carboxy-terminal processing
of the
large subunit of [NiFe] hydrogenases is associated with
the
incorporation of nickel into the protein, which is a prerequisite
for
translocation of the hydrogenase across the cytoplasmic membrane
(
25). For the dimeric [Fe] hydrogenase from
Desulfovibrio species,
it is likely that carboxy-terminal
processing of the large subunit
is not involved in the binding of the H
cluster to the protein,
since comparative analysis of gene sequence
indicates that the
monomeric [Fe] hydrogenases from clostridia which
are correctly
matured lack the corresponding sequence encoding the
precursor
C-terminal extension present in the large subunit of dimeric
hydrogenases
(
10,
16). Carboxy-terminal processing of
precursor proteins
destined to be exported to the periplasm or
transported into organelles
has been reported in a few cases (
7,
17). Several observations
indicate that the COOH-terminal
processing of the dimeric [Fe]
hydrogenase large subunit could be
involved in the export of the
protein to the periplasm: (i) cytoplasmic
monomeric [Fe] hydrogenases
from clostridia lack the gene sequence
coding for the C-terminal
extension found in the precursor form of the
large subunit of
dimeric [Fe] hydrogenase; (ii) the large subunit, in
contrast
to the small subunit, lacks an NH
2-terminal signal
peptide; and
(iii) the C-terminal extension has some characteristics in
common
with the import NH
2-terminal signal sequences which
target proteins
to inner compartments of eukaryotic organelles such as
the mitochondrial
matrix (
26,
30).
The NH
2-terminal sequences of the small subunits of
periplasmic or membrane-bound [Fe] or [NiFe] hydrogenases possess a
strictly
conserved motif (RRXFXK). This class of particular signal
sequences
has been extended to twin-arginine signal peptides that are
found
on precursors of bacterial periplasmic proteins binding various
types of redox cofactors (
3). Recently, a novel
Sec-independent
export system that may be used for the translocation of
these
folded proteins, including [NiFe] hydrogenase, has been
identified
in
E. coli (
4,
28). However, this
export system has not been
shown to function for the translocation of
[Fe] hydrogenase, since
such an enzyme is not present in this
bacterium. Since the catalytic
subunits of [Fe] hydrogenases (binding
the H cluster) and [NiFe]
hydrogenases (binding the Ni-Fe cluster)
lack an NH
2-terminal
signal sequence, it was notably
postulated that the single signal
peptide of the small subunit
containing the double-arginine motif
could mediate the export of both
subunits to the periplasm (
3,
40). The data reported here
indicate that the periplasmic [Fe]
hydrogenase from
Desulfovibrio species undergoes a C-terminal
processing of
its catalytic subunit, as is the case for [NiFe]-type
hydrogenase.
However, the biological significance of the two processes
appears to be
different: this processing is associated with the
incorporation of
nickel into the large subunit of [NiFe] hydrogenase,
whereas it might
be involved directly or indirectly in the export
of the enzyme to the
periplasm in the case of [Fe]
hydrogenase.
| |
ACKNOWLEDGMENTS |
We gratefully acknowledge J. Bonicel for mass spectrometric
analyses and N. Zylber for amino acid analyses. We are also indebted to
V. Méjean for critical reading of the manuscript.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Unité de
Bioénergétique et Ingéniérie des
Protéines, IBSM, CNRS, 31, Chemin Joseph Aiguier, 13402 Marseilles Cedex 20, France. Phone: 04 91 16 41 45. Fax: 04 91 16 45 78. E-mail: hatch{at}ibsm.cnrs-mrs.fr.
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REFERENCES |
| 1.
|
Adams, M. W. W.
1990.
The structure and mechanism of iron-hydrogenases.
Biochim. Biophys. Acta
1020:115-145[Medline].
|
| 2.
|
Albracht, S. P. J.
1994.
Nickel hydrogenases: in search for the active site.
Biochim. Biophys. Acta
1188:167-204[Medline].
|
| 3.
|
Berks, B. C.
1996.
A common export pathway for proteins binding complex redox cofactors?
Mol. Microbiol.
22:393-404[Medline].
|
| 4.
|
Bogsch, E. G.,
F. Sargent,
N. R. Stanley,
B. C. Berks,
C. Robinson, and T. Palmer.
1998.
An essential component of a novel bacterial protein export system with homologues in plastids and mitochondria.
J. Biol. Chem.
273:18003-18006[Abstract/Free Full Text].
|
| 5.
|
Casalot, L.,
E. C. Hatchikian,
N. Forget,
P. de Philip,
Z. Dermoun,
J.-P. Bélaich, and M. Rousset.
1998.
Molecular study and partial characterization of iron-only hydrogenase in Desulfovibrio fructosovorans.
Anaerobe
4:45-55.
|
| 6.
|
Devereux, R.,
S.-H. He,
C. L. Doyle,
S. Orkland,
D. A. Stahl,
J. LeGall, and W. B. Whitman.
1990.
Diversity and origin of Desulfovibrio species: phylogenetic definition of a family.
J. Bacteriol.
172:3609-3619[Abstract/Free Full Text].
|
| 7.
|
Diner, B. A.,
D. F. Ries,
B. N. Cohen, and J. G. Metz.
1988.
COOH-terminal processing of polypeptide D1 of the photosystem II reaction center of Scenedesmus obliquus is necessary for the assembly of the oxygen-evolving complex.
J. Biol. Chem.
263:8972-8980[Abstract/Free Full Text].
|
| 8.
|
Fauque, G.,
H. D. Peck, Jr.,
J. J. G. Moura,
B. H. Huynh,
Y. Berlier,
D. V. DerVartanian,
M. Teixeira,
A. E. Przybyla,
P. A. Lespinat,
I. Moura, and J. LeGall.
1988.
The three classes of hydrogenases from sulfate-reducing bacteria of the genus Desulfovibrio.
FEMS Microbiol. Rev.
54:299-344.
|
| 9.
|
Gollin, D. J.,
L. E. Mortenson, and R. L. Robson.
1992.
Carboxyl-terminal processing may be essential for production of active NiFe hydrogenase in Azotobacter vinelandii.
FEBS Lett.
309:371-375[Medline].
|
| 10.
|
Gorwa, M.-F.,
C. Croux, and P. Soucaille.
1996.
Molecular characterization and transcriptional analysis of the putative hydrogenase gene of Clostridium acetobutylicum ATCC 824.
J. Bacteriol.
178:2668-2675[Abstract/Free Full Text].
|
| 11.
|
Grande, H. J.,
W. R. Dunham,
B. Averill,
C. Van Dijk, and R. H. Sands.
1983.
Electron paramagnetic resonance and other properties of hydrogenases isolated from Desulfovibrio vulgaris (strain Hildenborough) and Megasphaera elsdenii.
Eur. J. Biochem.
136:201-207[Medline].
|
| 12.
|
Hagen, W. R.,
A. Van Berkel-Arts,
K. M. Kruse-Wolters,
G. Voordouw, and C. Veeger.
1986.
The iron-sulfur composition of the active site of hydrogenase from Desulfovibrio vulgaris (Hildenborough) deduced from its subunit structure and total iron-sulfur content.
FEBS Lett.
203:59-63.
|
| 13.
|
Hatchikian, E. C.,
N. Forget,
V. M. Fernandez,
R. Williams, and R. Cammack.
1992.
Further characterization of the [Fe] hydrogenase from Desulfovibrio desulfuricans ATCC 7757.
Eur. J. Biochem.
209:357-365[Medline].
|
| 14.
|
Higuchi, Y.,
T. Yagi, and N. Yasuoka.
1997.
Unusual ligand structure in Ni-Fe active center and a additional Mg site in hydrogenase revealed by high resolution X-ray structure analysis.
Structure
5:1671-1680[Medline].
|
| 15.
|
Magro, V.,
L. Pieulle,
N. Forget,
B. Guigliarelli,
Y. Petillot, and E. C. Hatchikian.
1997.
Further characterization of the two tetraheme cytochromes c3 from Desulfovibrio africanus: nucleotide sequences, EPR spectroscopy and biological activity.
Biochim. Biophys. Acta
1342:149-163[Medline].
|
| 16.
|
Meyer, J., and J. Gagnon.
1991.
Primary structure of hydrogenase I from Clostridium pasteurianum.
Biochemistry
30:9697-9704[Medline].
|
| 17.
|
Nagasawa, H.,
Y. Sakagami,
A. Suzuki,
H. Suzuki,
H. Hara, and Y. Hirota.
1989.
Determination of the cleavage site involved in C-terminal processing of penicillin-binding protein 3 of Escherichia coli.
J. Bacteriol.
171:5890-5893[Abstract/Free Full Text].
|
| 18.
|
Nicolet, Y.,
C. Piras,
P. Legrand,
E. C. Hatchikian, and J. C. Fontecilla-Camps.
1999.
Desulfovibrio desulfuricans iron hydrogenase: the structure shows unusual coordination to an active site Fe binuclear center.
Structure
7:13-23[Medline].
|
| 19.
|
Nivière, V.,
A. Bernadac,
N. Forget,
V. M. Fernandez, and E. C. Hatchikian.
1990.
Localization of hydrogenase in Desulfovibrio gigas cells.
Arch. Microbiol.
155:579-586.
|
| 20.
|
Nivière, V.,
S.-L. Wong, and G. Voordouw.
1992.
Site-directed mutagenesis of the hydrogenase signal peptide consensus box prevents export of a  -lactamase fusion protein.
J. Gen. Microbiol.
138:2173-2183[Medline].
|
| 21.
|
Odom, J. M., and H. D. Peck, Jr.
1984.
Hydrogenase, electron transfer proteins, and energy coupling in the sulfate-reducing bacteria Desulfovibrio.
Annu. Rev. Microbiol.
38:551-592[Medline].
|
| 22.
|
Patil, D. S.,
J. J. G. Moura,
S. H. He,
M. Teixeira,
B. C. Prickril,
D. V. DerVartanian,
H. D. Peck Jr,
J. LeGall, and B.-H. Huynh.
1988.
EPR-detectable redox centers of the periplasmic hydrogenase from Desulfovibrio vulgaris.
J. Biol. Chem.
263:18732-18738[Abstract/Free Full Text].
|
| 23.
|
Pierik, A. J.,
W. R. Hagen,
J. S. Redeker,
R. B. G. Wolbert,
M. Boresma,
M. F. J. M. Verhagen,
H. J. Grande,
C. Veeger,
P. H. A. Mutsaers,
R. H. Sands, and W. R. Dunham.
1992.
Redox properties of the iron-sulfur clusters in activated Fe-hydrogenase from Desulfovibrio vulgaris (Hildenborough).
Eur. J. Biochem.
209:63-72[Medline].
|
| 24.
|
Przybyla, A. E.,
J. Robbins,
N. Menon, and H. D. Peck, Jr.
1992.
Structure-function relationships among the nickel-containing hydrogenases.
FEMS Microbiol. Rev.
88:109-136.
|
| 25.
|
Rodrigue, A.,
D. H. Boxer,
M. A. Mandrand-Berthelot, and L.-F. Wu.
1996.
Requirement for nickel of the transmembrane translocation of NiFe-hydrogenase 2 in Escherichia coli.
FEBS Lett.
392:81-86[Medline].
|
| 26.
|
Roise, D.,
S. J. Horvath,
J. M. Tomich,
J. H. Richards, and G. Schatz.
1986.
A chemically synthesized pre-sequence of an imported mitochondrial protein can form an amphiphilic helix and perturb natural and artificial phospholipid bilayers.
EMBO J.
5:1327-1334[Medline].
|
| 27.
|
Rossman, R.,
M. Sauter,
F. Lottspeich, and A. Böck.
1994.
Maturation of the large subunit (HycE) of hydrogenase 3 of Escherichia coli requires nickel incorporation followed by C-terminal processing at Arg537.
Eur. J. Biochem.
220:377-384[Medline].
|
| 28.
|
Santini, C.-L.,
B. Ize,
A. Chanal,
M. Müller,
G. Giordano, and L.-F. Wu.
1998.
A novel Sec-independent periplasmic protein translocation pathway in Escherichia coli.
EMBO J.
17:101-112[Medline].
|
| 29.
|
Saunders, G. F.,
L. L. Campbell, and J. R. Postgate.
1964.
Base composition of deoxyribonucleic acid of sulfate-reducing bacteria deduced from buoyant density measurement in cesium chloride.
J. Bacteriol.
87:1073-1078[Abstract/Free Full Text].
|
| 30.
|
Schatz, G., and B. Dobberstein.
1996.
Common principles of protein translocation across membranes.
Science
271:1519-1526[Abstract].
|
| 31.
|
Subramani, S.
1996.
Convergence of model systems for peroxisome biogenesis.
Curr. Opin. Cell Biol.
8:513-518[Medline].
|
| 32.
|
Van der Westen, H.,
S. G. Mayhew, and C. Veeger.
1978.
Separation of hydrogenase from intact cells of Desulfovibrio vulgaris.
FEBS Lett.
86:122-126[Medline].
|
| 33.
|
Van Dongen, W.,
W. R. Hagen,
W. Van den Berg, and C. Veeger.
1988.
Evidence for an unusual mechanism of membrane translocation of the periplasmic hydrogenase of Desulfovibrio vulgaris (Hildenborough), as derived from expression in Escherichia coli.
FEMS Microbiol. Lett.
50:5-9.
|
| 34.
|
Volbeda, A.,
C. Piras,
M.-H. Charon,
E. C. Hatchikian,
M. Frey, and J. C. Fontecilla-Camps.
1995.
Crystal structure of the nickel-iron hydrogenase from Desulfovibrio gigas.
Nature
373:580-587[Medline].
|
| 35.
|
Volbeda, A.,
E. Garcin,
C. Piras,
A. I. de Lacey,
V. M. Fernandez,
E. C. Hatchikian,
M. Frey, and J. C. Fontecilla Camps.
1996.
Structure of the [NiFe] hydrogenase active site: evidence for biological uncommon Fe ligands.
J. Am. Chem. Soc.
118:12989-12996.
|
| 36.
|
von Heijne, G.
1986.
Mitochondrial targeting sequences may form amphiphilic helices.
EMBO J.
5:1335-1342[Medline].
|
| 37.
|
Voordouw, G., and S. Brenner.
1985.
Nucleotide sequence of the gene encoding the hydrogenase from Desulfovibrio vulgaris (Hildenborough).
Eur. J. Biochem.
148:515-520[Medline].
|
| 38.
|
Voordouw, G.,
J. D. Strang, and F. R. Wilson.
1989.
Organization of the genes encoding [Fe] hydrogenase in Desulfovibrio vulgaris subsp. oxamicus Monticello.
J. Bacteriol.
171:3881-3889[Abstract/Free Full Text].
|
| 39.
|
Voordouw, G.
1990.
Hydrogenase genes in Desulfovibrio, p. 37-51.
In
J. P. Bélaich, M. Bruschi, and J. L. Garcia (ed.), Microbiology and biochemistry of strict anaerobes involved in interspecies hydrogen transfer. Plenum Press, New York, N.Y.
|
| 40.
|
Voordouw, G.
1992.
Evolution of hydrogenase genes.
Adv. Inorg. Chem.
38:397-422.
|
Journal of Bacteriology, May 1999, p. 2947-2952, Vol. 181, No. 9
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Copyright © 1999, American Society for Microbiology. All rights reserved.
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