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Purification And Properties Of The Mercuric-ion-binding Protein MerP.

L. Sahlman, B. Jonsson
Published 1992 · Chemistry, Medicine

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The gene merP, coding for a mercuric-ion-binding periplasmic protein (P protein), was cloned into the expression vector pCA. In an Escherichia coli strain bearing the resulting plasmid, the P protein constitutes about 20% of total soluble protein. P protein was purified using ammonium sulfate precipitation and two chromatography steps. Typical yields were 20-30 mg from 7.5 l bacterial culture. The protein is a monomer with a molecular mass of 7500 Da. The periplasmic signal peptide was processed identically in both the recombinant and the wild-type proteins. CD spectra of both proteins were identical and indicated that the structure is highly ordered, containing approximately 80% alpha-helix. Purification in the presence of excess cysteine resulted in a form of the protein containing two reduced thiols, in agreement with the published sequence which has two cysteine residues. When cysteine was omitted from the purification buffers, no reduced thiol groups could be detected suggesting that the cysteine residues are oxidized. Both of these forms of the protein were found to bind approximately five Hg2+ ions/protein molecule in an apparently non-specific manner. However, in the presence of external thiol compounds, the protein with reduced thiols bound only one Hg2+ ion/protein molecule with an apparent Kd of 3.7 +/- 1.3 microM. Under these conditions, the protein with oxidized thiols did not bind Hg2+. The possible physiological role of this protein in Hg2+ detoxification is discussed.
This paper references
10.1073/PNAS.82.2.488
Rapid and efficient site-specific mutagenesis without phenotypic selection.
T. Kunkel (1985)
10.1128/JB.171.6.3458-3464.1989
Cloning and expression of Thiobacillus ferrooxidans mercury ion resistance genes in Escherichia coli.
T. Shiratori (1989)
10.1111/J.1574-6968.1990.TB04110.X
Bacterial periplasmic permeases belong to a family of transport proteins operating from Escherichia coli to human: Traffic ATPases.
G. F. Ames (1990)
10.1016/0968-0004(85)90069-6
Bacterial resistance to mercury — reductio ad absurdum?
N. Brown (1985)
10.1099/00221287-132-2-465
Transcriptional regulation of the mercury-resistance genes of transposon Tn501.
P. Lund (1986)
10.1016/S0168-6445(05)80008-7
Bacterial periplasmic permeases belong to a family of transport proteins operating from to human: Traffic ATPases
G. F. Ames (1990)
10.1073/pnas.74.12.5463
DNA sequencing with chain-terminating inhibitors.
F. Sanger (1977)
10.1128/JB.171.4.2222-2225.1989
Translation of merD in Tn21.
I. Lee (1989)
10.1146/ANNUREV.MI.32.100178.003225
Microbial transformations of metals.
A. Summers (1978)
10.1016/0378-1119(83)90180-4
Overexpression and purification of the sigma subunit of Escherichia coli RNA polymerase.
M. Gribskov (1983)
10.1016/0014-5793(92)81019-I
Carbonic anhydrase: From biochemistry and genetics to physiology and clinical medicine : Edited by F. Botrè, G. Gros and B.T. Storey; VCH Verlag; Weinheim, 1991; xvi + 467 pages. DM 186.00, £70.00. ISBN 352728365x
N. D. Carter (1992)
Improved method for the determination of blood glutathione.
E. Beutler (1963)
10.1128/JB.140.1.161-166.1979
Hypersensitivity to Hg2+ and hyperbinding activity associated with cloned fragments of the mercurial resistance operon of plasmid NR1.
H. Nakahara (1979)
10.1128/JB.162.2.773-776.1985
Some mercurial resistance plasmids from different incompatibility groups specify merR regulatory functions that both repress and induce the mer operon of plasmid R100.
T. Foster (1985)
10.1016/0378-1119(85)90134-9
Mercuric reductase structural genes from plasmid R100 and transposon Tn501: functional domains of the enzyme.
T. Misra (1985)
10.1146/ANNUREV.MI.40.100186.003135
Organization, expression, and evolution of genes for mercury resistance.
A. Summers (1986)
10.1016/0143-1471(80)90053-7
The Biogeochemistry of mercury in the environment
J. Nriagu (1979)
10.1016/0003-2697(87)90587-2
Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.
H. Schägger (1987)
10.1016/0378-1119(87)90047-3
Role of the merT and merP gene products of transposon Tn501 in the induction and expression of resistance to mercuric ions.
P. Lund (1987)
10.1002/PROT.340040407
Crystallographic studies of inhibitor binding sites in human carbonic anhydrase II: A pentacoordinated binding of the SCN− ion to the zinc at high pH
A. Eriksson (1988)
10.1128/JB.155.2.690-703.1983
Tn5 insertion mutations in the mercuric ion resistance genes derived from plasmid R100.
N. Nibhriain (1983)
10.1073/PNAS.81.19.5975
Mercuric ion-resistance operons of plasmid R100 and transposon Tn501: the beginning of the operon including the regulatory region and the first two structural genes.
T. Misra (1984)



This paper is referenced by
10.1016/S0168-6445(03)00046-9
Bacterial mercury resistance from atoms to ecosystems.
T. Barkay (2003)
10.1038/nsb0198-47
Solution structure of the fourth metal-binding domain from the Menkes copper-transporting ATPase
J. Gitschier (1998)
The Folding Energy Landscape of MerP
Ann-Christin Brorsson (2004)
10.1016/J.JTICE.2017.06.023
Copper, nickel, and zinc cations biosorption properties of Gram-positive and Gram-negative MerP mercury-resistance proteins
Yi-Huang Hsueh (2017)
10.1016/J.JMB.2004.03.022
Crystal structure of the oxidized form of the periplasmic mercury-binding protein MerP from Ralstonia metallidurans CH34.
L. Serre (2004)
10.1016/j.jhazmat.2008.04.079
Expressing a bacterial mercuric ion binding protein in plant for phytoremediation of heavy metals.
Ju-Liang Hsieh (2009)
The Folding Energy Landscape of The Folding Energy Landscape of The Folding Energy Landscape of The Folding Energy Landscape of MerP
Christin Brorsson (2004)
10.1046/J.1432-1327.1999.00680.X
Expression, purification and copper-binding studies of the first metal-binding domain of Menkes protein.
P. Y. Jensen (1999)
10.1016/S0065-2911(08)60158-7
Metal-microbe interactions: contemporary approaches.
T. J. Beveridge (1997)
10.1016/J.JMB.2006.01.090
GuHCl and NaCl-dependent hydrogen exchange in MerP reveals a well-defined core with an unusual exchange pattern.
Ann-Christin Brorsson (2006)
10.1002/9781119951438.EIBC0629
Structure of the Fourth Metal-Binding Domain from the Menkes Copper-Transporting ATPase
W. Fairbrother (2006)
10.1074/jbc.274.47.33320
Reactivity of the Two Essential Cysteine Residues of the Periplasmic Mercuric Ion-binding Protein, MerP*
J. Powlowski (1999)
10.1006/PREP.1997.0743
Effects of codon usage and vector-host combinations on the expression of spinach plastocyanin in Escherichia coli.
M. Ejdebäck (1997)
10.1002/ELAN.200604400
Protein-Based Capacitive Biosensors: a New Tool for Structure-Activity Relationship Studies
A. Mortari (2008)
10.1016/J.JMB.2004.05.003
The "two-state folder" MerP forms partially unfolded structures that show temperature dependent hydrogen exchange.
Ann-Christin Brorsson (2004)
10.1007/s00775-003-0495-y
Biophysical characterization of the MerP-like amino-terminal extension of the mercuric reductase from Ralstonia metallidurans CH34
Emmanuel Rossy (2003)
10.1016/j.febslet.2004.08.041
Is the cytoplasmic loop of MerT, the mercuric ion transport protein, involved in mercury transfer to the mercuric reductase?
Emmanuel Rossy (2004)
10.1038/KI.1995.52
Cytotoxicity of mercury compounds in LLC-PK1, MDCK and human proximal tubular cells.
H. H. Bohets (1995)
10.1007/978-90-481-3909-5_9
Heavy Metal Resistance in Pseudomonads
Esther Aguilar-Barajas (2010)
10.1007/s007920050097
The mer operon of the acidophilic bacterium Thiobacillus T3.2 diverges from its Thiobacillus ferrooxidans counterpart
A. Velasco (1999)
10.1111/j.1365-2958.1995.mmi_17010025.x
The role of cysteine residues in the transport of mercuric ions by the Tn501 MerT and MerP mercury‐resistance proteins
A. Morby (1995)
10.1006/BBRC.1999.0192
Roles of the four cysteine residues in the function of the integral inner membrane Hg2+-binding protein, MerC.
L. Sahlman (1999)
10.1074/jbc.272.47.29518
A Mercuric Ion Uptake Role for the Integral Inner Membrane Protein, MerC, Involved in Bacterial Mercuric Ion Resistance*
L. Sahlman (1997)
10.1271/bbb.70003
Overexpression of a Single Membrane Component from the Bacillus mer Operon Enhanced Mercury Resistance in an Escherichia coli Host
Ju-Liang Hsieh (2007)
10.1006/ABIO.1998.2689
Enzyme-complemented activatorsorbent assay (ECASA): genetic engineering for enzyme-linked immunosorbent assay-type mercuric ion detection.
J. Klein (1998)
10.1016/S0014-5793(97)00730-8
Remarkably slow folding of a small protein
G. Aronsson (1997)
10.1016/j.jhazmat.2020.122062
MerP/MerT-mediated mechanism: A different approach to mercury resistance and bioaccumulation by marine bacteria.
Jinlong Zhang (2020)
10.1007/978-3-319-20624-0_1
Metal Response in Cupriavidus metallidurans: Volume II: Insights into the Structure-Function Relationship of Proteins
G. Vandenbussche (2015)
10.1101/GR.196802
Metallochaperones and metal-transporting ATPases: a comparative analysis of sequences and structures.
F. Arnesano (2002)
10.1007/978-3-319-20624-0
Metal Response in Cupriavidus metallidurans: Volume II: Insights into the Structure-Function Relationship of Proteins
G. Vandenbussche (2015)
10.1016/S1367-5931(02)00314-9
Structural biology of metal-binding sequences.
S. Opella (2002)
10.1016/S0925-4439(98)00110-0
Characterisation of copper-binding to the second sub-domain of the Menkes protein ATPase (MNKr2).
M. D. Harrison (1999)
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