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Metal Uptake & Metallochaperones |
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Nickel Uptake
Trace
metals are essential for life. Organisms must obtain sufficient
quantities of trace metals for growth, while preventing the
accumulation of excess metal which can be toxic to the cell.
Our laboratory is interested in understanding the regulation
of cellular metal uptake, and we have begun our studies by
investigating Ni uptake in E. coli. Here, nickel concentrations
are controlled by transcriptional repression of the nickel-specific
ABC transporter. In the presence of excess intracellular nickel,
the nickel-dependent repressor NikR binds to an operator sequence,
turning off expression of the transporter and decreasing nickel
import. To gain insight into how nickel regulates NikR function,
we are determining crystal structures apo NikR, nickel-bound
NikR by itself, and the NikR:DNA complex. |
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Full-length apo-NikR |
Metal-binding domain
of NikR with Ni bound |
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Publications
Phillips CM, Schreiter ER, Stultz CM, and Drennan CL. (2010)
Structural Basis of Low Affinity Nickel Binding to the Nickel-Responsive
Transcription Factor NikR from Escherichia coli. Biochemistry.
49:7830-7838.
Schreiter, E.R., Sintchak, M.D., Guo, Y., Chivers, P.T., Sauer,
R.T., and Drennan, C.L. (2003), Crystal Structure
of the Nickel-Responsive Transcription Factor NikR,
Nature
Structural Biology.10:794-799.
Schreiter,
E.R, Wang, S. C., Zamble, D.B., Drennan, C.L. (2006)
NikR-operator complex structure and the mechanism of repressor activation
by metal ions, Proceedings
of the National Academy of Science U.S.A
103:13676-13681.
Phillips,
C.M., Schreiter, E.R., Guo, Y., Wang, S.C., Zamble, D.B., Drennan, C.L. (2008)
Structural Basis of the Metal Specificity for Nickel Regulatory Protein NikR, Biochemistry.
47(7):1938-1946. |
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Nickel Import in Escherichia Coli |
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Iron-Sulfur
Cluster Assembly
Iron sulfur clusters are ubiquitous cofactors involved in key cellular processes such as electron
transfer, gene regulation, and adenosyl- methionine-dependent radical chemistry. These clusters
can form spontaneously in vitro from aqueous iron (II) and sulfide; however, the in
vivo toxicity of these species requires strict regulation during cluster assembly. We are
interested in elucidating the structures and mechanisms of proteins involved in iron-sulfur
cluster assembly in Synechocystis. Thus far, we have determined the structure of the
essential cysteine desulfurase from Synechocystis to 1.8Å resolution. |
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| Cysteine Desulfurase |
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Publications
Tirupati, B., Vey , J. L., Drennan, C. L., Bollinger, Jr., J.M. (2004)
Kinetic and Structural Characterization of Slr0077/SufS, the Essential Cysteine
Desulfurase from Synechocystis sp. PCC 6803, Biochemistry.
43:12210-12219. |
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