British Heart Foundation Centre of Research Excellence
3-year Interdisciplinary PhD Studentship
Understanding the complex interaction between the redox state of PKG and PKA and their
regulation by cyclic nucleotides (Ref# 17/PE/NT02)
Professor Philip Eaton, Dr Gian de Nicola
Protein kinase G (PKG) and protein kinase A (PKA) are crucially important kinases in the cardiovascular system.
PKG is the end effector kinase that responds to elevations in cGMP which result from elevations in nitric oxide
(NO). PKA is the end effector kinase that responds to elevations in cAMP resulting from beta-adrenergic
signalling. In addition to being and being activated by cyclic nucleotides, PKGI and PKAI each also form
disulfide bonds during pro-oxidant stimuli.
This project is aimed at understanding the complex relationships between cyclic nucleotide binding to and the
redox state of these kinases. Such studies are important given their important roles in the cardiovascular system
and the regulation or dysregulation that oxidative stress can provide. These studies may provide new insight to
the role of oxidative stress in the maintenance of cardiovascular health and how this may go wrong during disease.
These studies will be challenging and utilise multiple, complementary techniques and approaches, including: Sitedirected mutagenesis, cell transfection, recombinant protein expression and purification, as well as binding assays
involving direct fluorescence polarisation, MicroScale Thermophoresis or surface plasmon resonance. Hands-on
practical experience in these techniques, as well as those relating to redox protein biochemistry (especially of
kinases) would be highly desirable.
Further reading includes:
1. Scotcher J, Prysyazhna O, Boguslavsky A, Kistamas K, Hadgraft K, Martin E D, Worthington J, Rudyk
O, Cutillas P. R, Cuello F, Shattock M J. , Marber M. S, Conte M R, Greenstein A, Greensmith D J,
Venetucci L, Timms J F, Eaton P (2016). Disulfide-activated protein kinase G Iα regulates diastolic
relaxation of the heart and fine-tunes the Frank-Starling response. Nature Communications (in press).
2. Burgoyne JR, Rudyk O, Cho H, Prysyazhna O, Evans R, Ng T, Schröder K, Brandes RP, Shah AM,
Eaton P (2015). Deficient angiogenesis in redox-dead Cys17Ser PKARI knock-in mice. Nature
Communications 6, article number:7920
3. De Nicola GF, Martin ED, Chaikuad A, Bassi R, Clark J, Martino L, Verma S, Sicard P, Tata R,
Atkinson RA, Knapp S, Conte MR, Marber MS (2013). Mechanism and consequence of the
autoactivation of p38α mitogen-activated protein kinase promoted by TAB1. Nat Struct Mol Biol.
20(10):1182-90. doi: 10.1038/nsmb.2668.
4. Prysyazhna O, Rudyk O, Eaton P (2012). Single atom substitution in mouse protein kinase G
eliminates oxidant sensing to cause hypertension. Nature Medicine. 18(2):286-90.
5. Burgoyne JR, Madhani M, Cuello F, Charles RL, Brennan JP, Schröder E, Browning DD and Eaton P
(2007). Cysteine redox sensor in PKGI enables oxidant-induced activation without nitric oxide
pathway. Science 317:1393-1397.
To apply for this application, please ensure to indicate reference #17/PE/NT02 in the
Project Title/Reference section on the Research Proposal section.