Characterisation of a novel cysteine/histidine-rich metal binding domain from Xenopus nuclear factor XNF7

Katherine L.B. Borden; Stephen R. Martin; Nicola J. O'Reilly; John M. Lally; Bramham A. Reddy; Laurence D. Etkin; Paul S. Freemont

doi:10.1016/0014-5793(93)80741-C

Characterisation of a novel cysteine/histidine-rich metal binding domain from Xenopus nuclear factor XNF7

Katherine L.B. Borden, Stephen R. Martin, Nicola J. O'Reilly, John M. Lally, Bramham A. Reddy, Laurence D. Etkin, Paul S. Freemont^*

^*Corresponding author for this work

Pharmacology

Research output: Contribution to journal › Article › peer-review

39 Scopus citations

Abstract

A 42 amino acid synthetic peptide corresponding to a newly defined cysteine/histidine-rich protein motif called B-box, from the Xenopus protein XNF7 has been characterised. The metal-binding stoichiometry and dissociation constant for zinc were determined by competition with the chromophoric chelator Br₂BAPTA, demonstrating that one zinc atom binds per molecule of peptide despite the presence of seven putative metal ligands, and represents the first application of this method to measuring zinc stoichiometry of proteins and/or peptides. Cobalt binding studies indicate that the motif binds zinc more tightly than cobalt, that cysteines are used as ligands and that the cation is co-ordinated tetrahedrally. Circular dichroism and NMR studies both indicate that the B-box peptide is structured only in the presence of zinc, copper and to a lesser extent cobalt.

Original language	English (US)
Pages (from-to)	255-260
Number of pages	6
Journal	FEBS Letters
Volume	335
Issue number	2
DOIs	https://doi.org/10.1016/0014-5793(93)80741-C
State	Published - Dec 6 1993

Funding

Ack~o~ledge~~rWs:e thank Darryl Pappin and Hans Hansenf or masss pectrometrayn alysiso f the B-box peptide.W e thankA ndrew Lanef or helpfuld iscussionos n them anuscripat nd Richard Newman and David Brown for readingt he final manuscriptT. his work was supportedb y the ImperialC ancerR esearchF und, Medical Research Councila ndN ationalS cienceF oundationD CB-900741t0o LDE and NC1 Training Grant No. NCI CA09299.S ome peptidesw ere syn-thesisedb y the SyntheticA ntigen Laboratorya t UTMDACC which is supportedb y NC1 CA16672.

Keywords

Metal binding
Protein motif
Zinc finger

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/0014-5793(93)80741-C

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title = "Characterisation of a novel cysteine/histidine-rich metal binding domain from Xenopus nuclear factor XNF7",

abstract = "A 42 amino acid synthetic peptide corresponding to a newly defined cysteine/histidine-rich protein motif called B-box, from the Xenopus protein XNF7 has been characterised. The metal-binding stoichiometry and dissociation constant for zinc were determined by competition with the chromophoric chelator Br2BAPTA, demonstrating that one zinc atom binds per molecule of peptide despite the presence of seven putative metal ligands, and represents the first application of this method to measuring zinc stoichiometry of proteins and/or peptides. Cobalt binding studies indicate that the motif binds zinc more tightly than cobalt, that cysteines are used as ligands and that the cation is co-ordinated tetrahedrally. Circular dichroism and NMR studies both indicate that the B-box peptide is structured only in the presence of zinc, copper and to a lesser extent cobalt.",

keywords = "Metal binding, Protein motif, Zinc finger",

author = "Borden, {Katherine L.B.} and Martin, {Stephen R.} and O'Reilly, {Nicola J.} and Lally, {John M.} and Reddy, {Bramham A.} and Etkin, {Laurence D.} and Freemont, {Paul S.}",

year = "1993",

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doi = "10.1016/0014-5793(93)80741-C",

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pages = "255--260",

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T1 - Characterisation of a novel cysteine/histidine-rich metal binding domain from Xenopus nuclear factor XNF7

AU - Borden, Katherine L.B.

AU - Martin, Stephen R.

AU - O'Reilly, Nicola J.

AU - Lally, John M.

AU - Reddy, Bramham A.

AU - Etkin, Laurence D.

AU - Freemont, Paul S.

PY - 1993/12/6

Y1 - 1993/12/6

N2 - A 42 amino acid synthetic peptide corresponding to a newly defined cysteine/histidine-rich protein motif called B-box, from the Xenopus protein XNF7 has been characterised. The metal-binding stoichiometry and dissociation constant for zinc were determined by competition with the chromophoric chelator Br2BAPTA, demonstrating that one zinc atom binds per molecule of peptide despite the presence of seven putative metal ligands, and represents the first application of this method to measuring zinc stoichiometry of proteins and/or peptides. Cobalt binding studies indicate that the motif binds zinc more tightly than cobalt, that cysteines are used as ligands and that the cation is co-ordinated tetrahedrally. Circular dichroism and NMR studies both indicate that the B-box peptide is structured only in the presence of zinc, copper and to a lesser extent cobalt.

AB - A 42 amino acid synthetic peptide corresponding to a newly defined cysteine/histidine-rich protein motif called B-box, from the Xenopus protein XNF7 has been characterised. The metal-binding stoichiometry and dissociation constant for zinc were determined by competition with the chromophoric chelator Br2BAPTA, demonstrating that one zinc atom binds per molecule of peptide despite the presence of seven putative metal ligands, and represents the first application of this method to measuring zinc stoichiometry of proteins and/or peptides. Cobalt binding studies indicate that the motif binds zinc more tightly than cobalt, that cysteines are used as ligands and that the cation is co-ordinated tetrahedrally. Circular dichroism and NMR studies both indicate that the B-box peptide is structured only in the presence of zinc, copper and to a lesser extent cobalt.

KW - Metal binding

KW - Protein motif

KW - Zinc finger

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ER -

Characterisation of a novel cysteine/histidine-rich metal binding domain from Xenopus nuclear factor XNF7

Abstract

Funding

Keywords

ASJC Scopus subject areas

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