Genetic and physiological characterization of new Escherichia coli mutants impaired in hydrogenase activity

Wu, Long-Fei; Mandrand‐Berthelot, Marie‐Andrée

doi:10.1016/s0300-9084(86)81081-1

Cited by 125 publications

(127 citation statements)

References 35 publications

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“…The addition of nickel to the growth medium specifically restores the intracellular nickel concentration and the hydrogenase activity in this mutant [15][16][17]. In this case, nickel is taken up through the nonspecific magnesium transport system.…”

Section: Effects Of Nickel On the Processing Of The Precursor Of The mentioning

confidence: 92%

“…atives of MC4100 (F-, araD139, A(argF-lac)U169, ptsF25, relA1, flbB5301, rpsL150, deoC1, rbsR, ~,-), are defective in the specific nickel transport system and therefore display a hydrogenase minus phenotype [15][16][17]. Addition of 300 ~M NiC12 to the growth medium results in restoration of the intracellular nickel concentration and hydrogenase activity in this mutant.…”

Section: Introductionmentioning

confidence: 98%

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Requirement for nickel of the transmembrane translocation of NiFe‐hydrogenase 2 in Escherichia coli

Rodrigue¹,

Boxer

Mandrand‐Berthelot³

et al. 1996

FEBS Letters

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The cellular location of membrane-bound NiFehydrogenase 2 (HYD2) from Escherichia coil was studied by immunobiot analysis and by activity staining. Treatment of spheroplasts with trypsin was able to release active HYD2 into the soluble fraction, indicating that HYD2 is attached to the periplasmic side of the cytoplasmic membrane and that HYD2 undergoes a trans-membrane translocation during its biosynthesis. By using a n/k mutant deficient in the high affinity specific nickel transport system, we show that the intracellniar availability of nickel is essential for the processing of the large subunit and for the transmembrane translocation of HYD2. We also demonstrate that the processing of the precursor, which is related with nickel incorporation, can occur in the membrane-depleted soluble fraction and that it is associated with the increase in resistance to proteolysis of the processed form of the large subunit. The mechanism of the transmembrane translocation of HYD2 is discussed.

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Section: Effects Of Nickel On the Processing Of The Precursor Of The mentioning

confidence: 92%

Section: Introductionmentioning

confidence: 98%

Requirement for nickel of the transmembrane translocation of NiFe‐hydrogenase 2 in Escherichia coli

Rodrigue¹,

Boxer

Mandrand‐Berthelot³

et al. 1996

FEBS Letters

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“…Mandrand-Berthelot et al showed that nik operon deletion led to the abolition of anaerobically-expressed [NiFe] hydrogenase activity in vivo [9]. This indicates that E. coli requires active nickel transport to synthesize these enzymes.…”

Section: The Role Of Histidine 416 In Nickel Uptake Is Reflected By Hmentioning

confidence: 99%

“…Once in E. coli's periplasm, nickel import through the inner membrane proceeds via a unique ATP-Binding Cassette (ABC)-type transporter called Nik-ABCDE encoded by the nik operon [7]. It is suggested to be a hydrogenase-specific nickel transporter [8] and insertion mutations in the nik operon completely abolish [NiFe] hydrogenase activity [9]. Its expression is under the positive control of the oxygen sensor Fumarate Nitrate reductase Regulator (FNR) [10], which forms a DNA-binding dimer during anaerobic growth, and is negatively controlled by the transcriptional metalloregulator NikR in the presence of excess nickel [11].…”

Section: Introductionmentioning

confidence: 99%

Histidine 416 of the periplasmic binding protein NikA is essential for nickel uptake in Escherichia coli

Cavazza¹,

Martin²,

Laffly³

et al. 2011

FEBS Letters

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Edited by Stuart Ferguson Keywords:Nickel uptake Escherichia coli NikA Periplasmic binding protein [NiFe] hydrogenase a b s t r a c t Escherichia coli require nickel for the synthesis of [NiFe] hydrogenases under anaerobic growth conditions. Nickel import depends on the specific ABC-transporter NikABCDE encoded by the nik operon, which deletion causes the complete abolition of hydrogenase activity. We have previously postulated that the periplasmic binding protein NikA binds a natural metallophore containing three carboxylate functions that coordinate a Ni(II) ion, the fourth ligand being His416, the only direct metal-protein contact, completing a square-planar coordination for the metal. The crystal structure of the H416I mutant showed no electron density corresponding to a metal-chelator complex. In vivo experiments indicate that the mutation causes a significant decrease in nickel uptake and hydrogenase activity. These results confirm the essential role of His416 in nickel transport by NikA.

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“…Mutation of the nik operon results in a hydrogenase-negative phenotype that can be specifically restored by addition of nickel salt to the growth medium (Wu and Mandrand-Berthelot, 1986;Wu et al, 1989).…”

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confidence: 99%

Purification and Characterization of the Periplasmic Nickel‐Binding Protein NikA of Escherichia coli K12

Pina

Navarro

Mcwalter

et al. 1995

European Journal of Biochemistry

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The nik operon of Escherichia coli encodes a periplasmic binding-protein-dependent transport system specific for nickel. In this report, we describe the overproduction of the periplasmic nickel-binding protein NikA by cloning the nikA gene into an overexpression vector, pREl. NikA was purified free of nickel to near homogeneity from the periplasm by hydrophobic and ion-exchange chromatography. N-terminal amino acid sequencing confirmed that the leader peptide of NikA had been removed. The nickel-binding properties of the protein has been studied by monitoring the quenching of intrinsic protein fluorescence.NikA binds one atom of nickellmolecule of protein with a dissociation constant (Kd) of less than 0.1 pM.Other metals (cobalt, copper, iron) are bound at least 10-fold less tightly. The high specificity for Ni2+ is also demonstrated by high-performance immobilized-metal-ion affinity chromatography. Biosynthesis of NikA occurred only under anaerobic conditions and was dependent on the general anaerobic regulator FNR. It was repressed by the presence of 250 pM NiZ+ in the growth medium and was not affected by either 30 mM formate or 100 mM nitrate. Anaerobically grown wild-type strain MC4100 contains about 23000 molecules of NiWcell. In addition to the effect on nickel transport, nikA mutation affects also the nickel sensing in Tar-dependent repellent chemotaxis.Keywords. nikA gene ; nickel binding ; periplasmic protein ; transport; overproduction.Bacterial cold-osmotic-shock-sensitive transport systems are complex permeases composed of a periplasmic substrate-binding receptor and a membrane-bound complex containing 2 -4 proteins (Ames, 1986). These permeases are energized by the hydrolysis of ATP, mediated by one or two of the proteins in the complex. The periplasmic substrate-binding proteins are the primary receptors for the transport systems specific for different small molecules such as sugars, amino acids, peptides and inorganic ions. In some cases, they also serve as the primary receptors for bacterial chemotaxis (Bourret et al., 1991;Furlong, 1987;Macnab, 1987). For example, in addition to its role in transport, the periplasmic maltose-binding protein, MBP, encoded by ma1E functions as the primary maltose chemoreceptor (Koiwai and Hayashi, 1979;Richarme, 1982). Generation of the chemotactic response to maltose requires a second protein, the chemotactic signal transducer Tar, which is also required for the chemotactic response to aspartate and to two repellents Ni2+ and Co2+ (Springer et al., 1977;Tso and Adler, 1974 Nickel has long been known as a heavy metal toxic to both eukaryotic and prokaryotic organisms (Barbich and Stotzky, 1983 ;Brown and Sunderman, 1988). Epidemiological studies have identified nickel as potentially carcinogenic and allergenic to humans (Dool et al., 1977; MennC et al., 1989). We demonstrated recently that nickel has an antagonistic effect on the fermentative growth of Escherichia coli . In E.coli the only known nickel-containing enzymes are the three hydrogenase isoenzymes whic...

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Genetic and physiological characterization of new Escherichia coli mutants impaired in hydrogenase activity

Cited by 125 publications

References 35 publications

Requirement for nickel of the transmembrane translocation of NiFe‐hydrogenase 2 in Escherichia coli

Requirement for nickel of the transmembrane translocation of NiFe‐hydrogenase 2 in Escherichia coli

Histidine 416 of the periplasmic binding protein NikA is essential for nickel uptake in Escherichia coli

Purification and Characterization of the Periplasmic Nickel‐Binding Protein NikA of Escherichia coli K12

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