TRACKING ATP-DEPENDENT PROTEIN DYNAMICS

Adenosine triphosphate (ATP) drives many critical cellular processes. We will use laser-induced release from a caged, inactive form of ATP followed by short X-ray pulses at the dedicated time-resolved X-ray solution scattering (TR-XSS) beamline ID09 at ESRF-EBS to study protein conformational change in real time. We aim to identify fast-forming ATP-binding states in both bacterial and eukaryotic ATP-driven Ca2+ transport, and to monitor how the lipidic environment affect functional structural rearrangements. We will use the same pump-and-probe methodology to track catalysis of nucleotide interconversion, which is central to cellular hemostasis. Here, we aim to determine kinetics and structural dynamics of fast-forming intermediates associated with ATP binding. The results will provide better understanding of lipid-induced membrane transport regulation, cooperative protein dynamics, and significantly increase the number of protein targets for similar time-resolved X-ray studies.