Advances in imaging of understudied ions in signaling: A focus on magnesium.

The study of metal ions in the context of cell signaling has historically focused mainly on Ca2+, the second messenger par excellence. But recent studies support an emerging paradigm in which other metals, including magnesium and d-block metals, play a role in signal transduction as well. Armed with the right indicators, fluorescence microscopy offers a unique combination of spatial and temporal resolution perfectly suited to reveal metal transients in real time, while also helping identify possible sources of ion mobilization and molecular targets. With a focus on Mg2+, we highlight recent advancements in the development of molecular indicators and imaging strategies for the study of metal ions in signaling. We discuss remaining conceptual and technical challenges in the field, and we illustrate through the case of Mg2+ how the study of nontraditional ions in signaling is inspiring technological developments applicable more broadly to the study of metals in biology.

Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals.

Innovative new crystallographic methods are facilitating structural studies from ever smaller crystals of biological macromolecules. In particular, serial X-ray crystallography and microcrystal electron diffraction (MicroED) have emerged as useful methods for obtaining structural information from crystals on the nanometre to micrometre scale. Despite the utility of these methods, their implementation can often be difficult, as they present many challenges that are not encountered in traditional macromolecular crystallography experiments. Here, XFEL serial crystallography experiments and MicroED experiments using batch-grown microcrystals of the enzyme cyclophilin A are described. The results provide a roadmap for researchers hoping to design macromolecular microcrystallography experiments, and they highlight the strengths and weaknesses of the two methods. Specifically, we focus on how the different physical conditions imposed by the sample-preparation and delivery methods required for each type of experiment affect the crystal structure of the enzyme.