cecilia piergentili cv
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Cecilia Piergentili1 Finney Court, Finney Terrace, DH1 1RX Durham (UK); [email protected];
Tel. +44 7547285892
PROFESSIONAL EXPERIENCE
October 2015 – ongoing ● School of Biological and Biomedical Sciences
The project aims to assist in the analysis of metal contents of biological samples by inductively
coupled plasma mass spectrometry (ICP-MS) in a collaborative project entitled: ”Unravelling and
engineering the role of trace metals on recombinant therapeutic protein synthesis and heterogeneity
from Chinese hamster ovary cells” in collaboration with the University of Kent and Lonza.
Sample preparation and quantification of elemental contents (ICP-MS)
Maintenance of analytical facilities
Training to academic and commercial users
Writing of progress reports for industrial partners and research publications
Networking with industrial and academic researchers in support of the delivery of project.
Aware of University policies on Data Protection and Health and Safety.
November 2011 – October 2015 ● School of Biological and Biomedical Sciences
Thesis title: “Metal-sensing in Salmonella typhimurium : A model for targeting a network that
differentiates metals”
Researcher supervisors: Prof. Nigel Robinson, Dr. Ehmke Pohl
Funded by BBSRC and Procter and Gamble
SUMMARY • PhD student in Protein Chemistry
• Available from October 2015
• Lateral / creative thinking
• Experienced in working in an interdisciplinary environment
EDUCATION
PhD in BIOLOGICAL SCIENCES DURHAM UNIVERSITY
Graduate Research Assistant in BIOLOGICAL SCIENCES DURHAM UNIVERSITY
Research highlights:
Metals are essential for biology and are required for proper function of around half of all enzymes.
My PhD work focussed on dissecting how levels of these metals are selectively balanced and
managed by regulatory proteins within a bacterial system. I investigated factors determining selective
responses of DNA-binding metal-sensors, including analysis of the formaldehyde sensor FrmR.
Salmonella FrmR, a member of the RcnR/CsoR family of metalloregulators, has been characterised
during the course of this work and was coincidentally confirmed to bind specifically to frmA operon,
which encodes a putative Zn(II)-requiring class III alcohol dehydrogenase. FrmR shares a high degree
of similarity with Ni(II)/Co(II)-sensing RcnR, in particular conserving two residues of a so-called
WXYZ motif required to detect metals. Metal-binding properties of FrmR were therefore extensively
investigated in vitro and its ability, or otherwise, to respond to metals explored in vivo.
FrmR binds Zn(II), Cu(I), Co(II) and Ni(II), adopting different geometries, and always involving a
mercapto group from the only cysteine residue (Cys35). Moreover, KZn(II)FrmR is in the range of affinity
found for other zinc sensors.
Since FrmR fails to sense metals in cells, where only formaldehyde is detected, questions about which
parameters are required in metal regulation in Salmonella and, in general, in bacteria were
investigated. A single-point mutation (Glu64 His) allows FrmR to sense cellular zinc and cobalt.
FrmR and E64HFrmR have been consequently used as a case of study to test hypotheses about the
mechanisms determining which metals are detected by a given sensor in cells.
In addition, the ability of FrmR to detect cellular formaldehyde has been investigated, and a reaction
mechanism tested by site-directed mutagenesis in vitro. Salmonella Ni(II)/Co(II)-sensing RcnR has
been characterised, and employed to test the specificity of formaldehyde responsiveness of FrmR. By
a single point-mutation (Ser2 Pro), the Ni(II)/Co(II)-sensing RcnR has been successfully switched
to a formaldehyde sensor in vitro, further endorsing the proposed mechanism.
Investigation of FrmR structure has been pursued by producing crystals of apo- and Zn(II)-protein
forms, which were then analysed at the Diamond Light Source. Investigation of FrmR structure has
been pursued by producing crystals of apo- and Zn(II)-protein forms, which were then analysed at the
Diamond Light Source.
PhD results were incorporated into a research paper (see below).
Publications:
“Generating a Metal-Responsive Transcriptional Regulator to Test What Confers Metal-Sensing in Cells”Deenah Osman*, Cecilia Piergentili*, Junjun Chen, Buddhapriya Chakrabarti, Andrew W Foster, Elena Lurie-Luke, Thomas G Huggins, and Nigel J Robinson
* Joint first author
Research supervisor: Dr. Dan Lloyd
Research highlights:
• Production of therapeutic protein from initial bio-informatic analysis and genetic design stage
• Transfection, expression in host system, protein purification and characterization, and
functional analysis.
November 2009-October 2011, Dept. of Chemistry
• 110/110 cum laude, equivalent to a 1.1 honour degree
Thesis title: “Thiosemicarbazones and their Cu(II) and Ni(II) complexes as potential novel
topoisomerase IIA inhibitors: Synthesis, characterization and biological studies”
Research supervisors: Prof. Giorgio Pelosi, Dr. Franco Bisceglie
Research period: March 2011-October 2011
Research highlights:
Synthesis of different thiosemicarbazones followed by a characterization through standard procedures
biological tests. Our attention was mainly devoted to the anticancer activity of a series of compounds
based on thiosemicarbazone complexes of copper and nickel. The biological target was the inhibition
of Topoisomerase II. More in detail, I tested the action mechanisms of those metal complexes with
particular emphasis on the role played by the substitutions made on the ligand on the biological
activity of the complexes.
October 2006-October 2009, Dept. of Chemistry
• 106/110, equivalent to a 2.1 degree
Thesis title: “Synthesis, characterization and stability test of inorganic self-assembled supramolecules
known as metallacrowns”
Research supervisor: Dr. Matteo Tegoni
Research period: April 2009-September 2009
Summer school in BIOTECHNOLOGY UNIVERSITY OF KENT
July 2011- August 2011, School of Biosciences
MSc in CHEMISTRY PARMA UNIVERSITY (Italy)
BSc in CHEMISTRY PARMA UNIVERSITY (Italy)
Research highlights:
My work involved the synthesis and characterization of inorganic self-assembled supramolecules
known as metallacrowns. I investigated metallacrowns stability in presence of competing ligands and
managed study of core metal substitution performed by UV-visible titrations and data treatment.
The results achieved were fundamental to demonstrate the functional analogies between
metallacrowns and crown ethers, and my participation to these studies was acknowledged in the
related publication (M.Tegoni, V.L.Pecoraro, et al., Inorg. Chem. 49 (2010) 5190)
- Cloning/Overexpression/Purification of native, recombinants proteins. Particular experience
in purifying DNA-binding, metal-sensing transcriptional regulators.
Molecular cloning, site-directed mutagenesis, SDS-PAGE analysis, FPLC (AKTA Purification
System), metal-affinity, heparin-affinity, ion exchange, size-exclusion chromatography
techniques
- In vitro Protein Characterisation
UV-VIS, fluorescence spectroscopy, gel filtration chromatography, DNA binding assay (EMSA,
Fluorescence anisotropy spectroscopy), metalloprotein-chelator competition experiments (for
determination of protein-metal binding affinities)
- Elemental composition analysis of purified, recombinant metalloproteins and other biological
samples
ICP-MS techniques
- Preparation/Characterisation of purified, reduced recombinant metalloprotein (and small
molecule) samples in strict anaerobic environments
Anaerobic glovebox
- Structural characterisation of proteins
Protein crystallography, CD Spectroscopy
- Acquisition of skills in the synthesis of organic ligands with protection-activation strategies
under controlled conditions
Schlenk glassware, purified and dried solvents, and vacuum lines
PROCESSES AND TECHNOLOGIES
- Trained in the used of the principal characterization techniques of organic and inorganic
compounds
NMR, IR, MS, etc.
- Languages:
English (full professional proficiency)
Italian (native proficiency)
- Computer literate (all MS Office suite, DynaFit, SigmaPlot).
- Experienced in planning, designing and conducting experiments, analysing data and
communicating results to colleagues and collaborators via meetings and conferences.
- Contributor to the planning, editing and submission of research papers.
- Mentoring: experienced in mentoring colleagues, sharing technical expertise and delivering
practical classes to undergraduate students.
- Microbiology lab experience (Laboratory Biosafety Level Criteria: BSL-1 and BSL-2).
- Ability to replicate technical procedures accurately.
- Self-disciplined and motivated when working in isolation.
- Good team worker when undertaking research with academic colleagues.
- Perseverant, able to overcome difficulties.
- Willing to learn new techniques.
- As a Celebrating Science 2014 Edition demonstrator, I run chemistry themed activities with
the aim to inspire and educate local secondary school students about science.
- Member of a running club; I have run the Dublin Half Marathon 2014 and the Edinburgh Half
Marathon 2015 and I am looking for the next challenge. I have also taken part in the 5k Race
for Life event in Durham and helped to raise money to fund Cancer Research UK.
ADDITIONAL SKILLS
HOBBIES AND INTERESTS
Prof. Nigel J. Robinson
Professor in the School of Biological and Biomedical Sciences,
Dept. of Chemistry,
Durham University,
South Road, Durham
DH1 3LE,
United Kingdom
+44(0)1913342143
Dr. Ehmke Pohl
Reader in the School of Biological and Biomedical Sciences,
Dept. of Chemistry,
Durham University,
South Road, Durham
DH1 3LE,
United Kingdom
+44(0)1913343619
Dr. Deenah Osman
Research Associate in the School of Biological and Biomedical Sciences,
Dept. of Chemistry,
Durham University,
South Road, Durham
DH1 3LE,
United Kingdom
+44(0)1913342143
REFERENCES