Synthetic metallochaperone ZMC1 rescues mutant p53 conformation by transporting zinc into cells as an ionophore

Adam R Blanden; Xin Yu; Aaron J Wolfe; John A Gilleran; David J Augeri; Ryan S O'Dell; Eric C Olson; S David Kimball; Thomas J Emge; Liviu Movileanu; Darren R Carpizo; Stewart N Loh

doi:10.1124/mol.114.097550

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Synthetic metallochaperone ZMC1 rescues mutant p53 conformation by transporting zinc into cells as an ionophore

Journal article

Open access

Peer reviewed

Synthetic metallochaperone ZMC1 rescues mutant p53 conformation by transporting zinc into cells as an ionophore

Adam R Blanden, Xin Yu, Aaron J Wolfe, John A Gilleran, David J Augeri, Ryan S O'Dell, Eric C Olson, S David Kimball, Thomas J Emge, Liviu Movileanu, …

Molecular pharmacology, Vol.87(5), pp.825-831

05/2015

DOI: https://doi.org/10.1124/mol.114.097550

PMCID: PMC4407733

PMID: 25710967

Abstract

Cell Line

Ionophores - metabolism

Zinc - metabolism

Humans

Tumor Suppressor Protein p53 - metabolism

Mutation - genetics

Tumor Suppressor Protein p53 - genetics

Carrier Proteins - metabolism

HEK293 Cells

Protein Conformation

Biological Transport - physiology

Cell Membrane - metabolism

Metallochaperones - metabolism

Binding Sites

p53 is a Zn(2+)-dependent tumor suppressor inactivated in >50% of human cancers. The most common mutation, R175H, inactivates p53 by reducing its affinity for the essential zinc ion, leaving the mutant protein unable to bind the metal in the low [Zn(2+)]free environment of the cell. The exploratory cancer drug zinc metallochaperone-1 (ZMC1) was previously demonstrated to reactivate this and other Zn(2+)-binding mutants by binding Zn(2+) and buffering it to a level such that Zn(2+) can repopulate the defective binding site, but how it accomplishes this in the context of living cells and organisms is unclear. In this study, we demonstrated that ZMC1 increases intracellular [Zn(2+)]free by functioning as a Zn(2+) ionophore, binding Zn(2+) in the extracellular environment, diffusing across the plasma membrane, and releasing it intracellularly. It raises intracellular [Zn(2+)]free in cancer (TOV112D) and noncancer human embryonic kidney cell line 293 to 15.8 and 18.1 nM, respectively, with half-times of 2-3 minutes. These [Zn(2+)]free levels are predicted to result in ∼90% saturation of p53-R175H, thus accounting for its observed reactivation. This mechanism is supported by the X-ray crystal structure of the [Zn(ZMC1)2] complex, which demonstrates structural and chemical features consistent with those of known metal ionophores. These findings provide a physical mechanism linking zinc metallochaperone-1 in both in vitro and in vivo activities and define the remaining critical parameter necessary for developing synthetic metallochaperones for clinical use.

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Title: Synthetic metallochaperone ZMC1 rescues mutant p53 conformation by transporting zinc into cells as an ionophore
Creators: Adam R Blanden - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
Xin Yu - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
Aaron J Wolfe - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
John A Gilleran - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
David J Augeri - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
Ryan S O'Dell - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
Eric C Olson - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
S David Kimball - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
Thomas J Emge - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
Liviu Movileanu - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
Darren R Carpizo - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.)
Stewart N Loh - Department of Biochemistry and Molecular Biology (A.R.B.,S.N.L.) and Department of Neuroscience and Physiology (R.S.O., E.C.O.), State University of New York Upstate Medical University, Syracuse, New York; Rutgers Cancer Institute of New Jersey (X.Y., D.R.C.), Department of Surgery, Rutgers Robert Wood Johnson Medical School (X.Y., D.R.C.), Office of Translational Sciences (J.A.G., D.J.A., S.D.K.), and Department of Chemistry and Chemical Biology (T.J.E.), Rutgers University, New Brunswick, New Jersey; and Department of Physics, Syracuse University, Syracuse, New York (A.J.W., L.M.) lohs@upstate.edu
Publication Details: Molecular pharmacology, Vol.87(5), pp.825-831
Date published: 05/2015
Grant note: R01 GM088403 / NIGMS NIH HHS R01-GM088403 / NIGMS NIH HHS K08 CA172676 / NCI NIH HHS K08-CA172676-02 / NCI NIH HHS P30 CA072720 / NCI NIH HHS F30 GM113299 / NIGMS NIH HHS
Academic Unit: Chemistry and Chemical Biology (SAS); Medicinal Chemistry (EMSOP); Rutgers Translational Sciences; Office of CAOO; Office of Research and Economic Development
Language: English
Resource Type: Journal article
Identifiers: 991031654029304646