Metal binding and folding properties of a minimalist Cys2His2 zinc finger peptide.

Michael, Scott F.; Kilfoil, Valda J.; Schmidt, Michael; Amann, Barbara; Berg, Jeremy M.

doi:10.1073/pnas.89.11.4796

Cited by 162 publications

(186 citation statements)

References 26 publications

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“…The most dramatic demonstration of the plasticity of the domain comes from the work of Berg and coworkers on a "minimalist" zinc finger called MZF (Michael et al, 1992). The MZF sequence preserves the two conserved aromatics at positions 104 and 113 (ADRlb numbering scheme), the conserved leucine at position 119, and the cysteine and histidine ligands, with the remaining residues being alanine (with lysine at positions 103, 116, and 128 to promote solubility). '…”

Section: Discussionmentioning

confidence: 99%

Structures of DNA‐binding mutant zinc finger domains: Implications for DNA binding

Hoffman¹,

Horvath²,

Klevit³

1993

Protein Science

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Studies of Cys,-His2 zinc finger domains have revealed that the structures of individual finger domains in solution determined by NMR spectroscopy are strikingly similar to the structure of fingers bound to DNA determined by X-ray diffraction. Therefore, detailed structural analyses of single finger domains that contain amino acid substitutions known to affect DNA binding in the whole protein can yield information concerning the structural ramifications of such mutations. We have used this approach to study two mutants in the N-terminal finger domain of ADRl, a yeast transcription factor that contains two Cys2-His2 zinc finger sequences spanning residues 102-159. Two point mutants at position 118 in the N-terminal zinc finger (ADRlb: 102-130) that adversely affect the DNA-binding activity of ADRl have previously been identified: H118A and H118Y. The structures of wild-type ADRlb and the two mutant zinc finger domains were determined using two-dimensional nuclear magnetic resonance spectroscopy and distance geometry and were refined using a complete relaxation matrix method approach (REPENT) to improve agreement between the models and the nuclear Overhauser effect spectroscopy data from which they were generated. The molecular architecture of the refined wild-type ADRlb domain is presented in detail. Comparisons of wild-type ADRlb and the two mutants revealed that neither mutation causes a significant structural perturbation. The structures indicate that the DNA binding properties of the His 118 mutants are dependent on the identity of the side chain at position 118, which has been postulated to make a direct DNA contact in the wild-type ADRl protein. The results suggest that the identity of the side chain at the middle DNA contact position in Cys2-His2 zinc fingers may be changed with impunity regarding the domain structure and can affect the affinity of the protein-DNA interaction.Keywords: DNA binding; molecular architecture; NMR; protein-DNA interaction; zinc finger Analysis of the deduced amino acid sequence of the Xenprotein (Miller et al., 1985) and that zinc was required for opus transcription factor TFIIIA in the mid-1980s revealed sequence specific DNA binding (Hanas et al., 1983). that it contains nine tandem repeats of the consensus seThese and other findings were synthesized in the zinc fin- quence (Y ,F)-X-C-X2,4-C-X3-F-X5-L-X2-H-X3.5-H-X2-6ger hypothesis that proposed each of the consensus re- (Brown et al., 1985;Miller et al., 1985). It was further peats binds a zinc ion through the conserved cysteine and noted that when purified in the absence of metal-chelathistidine residues to form a stable structural domain that ing agents, 7-1 1 moles of zinc co-purified per mole of is responsible for the DNA-binding activity of the transcription factor (Brown et al., 1985;Miller et al., 1985). ~~Since that time, a large number of Cysz-His2 zinc finger finger sequences (Berg, 1990).H-bonds, hydrogen bonds; COSY, correlation spectroscopy; RELAY, relayed coherence transfer spectroscopy; TOCSY, total co...

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Section: Discussionmentioning

confidence: 99%

Structures of DNA‐binding mutant zinc finger domains: Implications for DNA binding

Hoffman¹,

Horvath²,

Klevit³

1993

Protein Science

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“…Hydrophobic residues, which are also well conserved (although their spacing within the domain changes), provide additional stability by packing into the core of the fold (Michael et al, 1992;Miller et al, 1985;Page et al, 1987;Shi and Berg, 1995;Weiss and Keutmann, 1990).…”

Section: Structurementioning

confidence: 99%

The Protein-Binding Potential of C2H2 Zinc Finger Domains

Brayer

Kulshreshtha

Segal

2008

Cell Biochem Biophys

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“…Dissociation constant of Pb-F3-The binding of Co 2+ to F3 was followed by the change of the absorbance spectrum at 650 nm during the titration of apo-F3 with Co 2+ in anaerobic 20 mM Tris/Cl, 100 mM NaCl, pH 7.2 [43]. The apparent dissociation constant of Co(II)-F3 was calculated directly from the spectrophotometric titration as previously described, using the expression [43]: (1) where A max is the final absorbance at 650 nm and A represents any intermediate absorbance during the titration corresponding to free [Co 2+ ].…”

Section: Co 2+ Pb 2+ Titrations Of Fingermentioning

confidence: 99%

“…The apparent dissociation constant of Co(II)-F3 was calculated directly from the spectrophotometric titration as previously described, using the expression [43]: (1) where A max is the final absorbance at 650 nm and A represents any intermediate absorbance during the titration corresponding to free [Co 2+ ]. Then, the apparent dissociation constant of Pb-F3 was determined by competitive displacement of Co 2+ from Co-F3 with Pb 2+ , observing the decrease in the absorbance (A) of the 650 nm band of Co(II)-F3.…”

Section: Co 2+ Pb 2+ Titrations Of Fingermentioning

confidence: 99%

Zn-, Cd-, and Pb-transcription factor IIIA: properties, DNA binding, and comparison with TFIIIA-finger 3 metal complexes

Huang

Krepkiy

et al. 2004

Journal of Inorganic Biochemistry

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Properties of the metal ion binding sites of Zn-transcription factor IIIA (TFIIIA) were investigated to understand the potential of this type of zinc finger to undergo reactions that remove Zn 2+ from the protein. Zn-TFIIIA was purified from E. coli containing the cloned sequence for Xenopus laevis oocyte TFIIIA and its stoichiometry of bound Zn 2+ was shown to depend on the details of the isolation process. The average dissociation constant of Zn 2+ in Zn-TFIIIIA was 10 −7 . The dissociation constant for Zn-F3, the third finger from the N-terminus of TFIIIA, was 1.0 × 10 −8 . The reactivity of Zn-TFIIIA with a series of metal binding ligands, including 2-carboxy-2′-hydroxy-5′-sulfoformazylbenzene (zincon), 4-(2-pyridylazo)-resorcinol (PAR), and 3-ethoxy-2-oxo-butyraldehyde-bis-(N 4 -dimethylthiosemicarbazone) (H 2 KTSM 2 ) revealed similar kinetics. The reactivity of PAR with Zn-TFIIIA declined substantially when the protein was bound to the internal control region (ICR) of the 5S ribosomal DNA. Both Cd 2+ and Pb 2+ disrupt TFIIIA binding to its cognate DNA sequence. The Pb 2+ dissociation constant of Pb-F3 was measured as 2.5 × 10 −8 . According to NMR spectroscopy, F3 does not fold into a regular conformation in the presence of Pb 2+ .

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Metal binding and folding properties of a minimalist Cys2His2 zinc finger peptide.

Cited by 162 publications

References 26 publications

Structures of DNA‐binding mutant zinc finger domains: Implications for DNA binding

Structures of DNA‐binding mutant zinc finger domains: Implications for DNA binding

The Protein-Binding Potential of C2H2 Zinc Finger Domains

Zn-, Cd-, and Pb-transcription factor IIIA: properties, DNA binding, and comparison with TFIIIA-finger 3 metal complexes

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