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Instruments / Technical Area	Publication Details	Published
Krios II-Titan Krios II at Diamond	The role of macromolecular condensates in the regulation of intracellular calcium transport for coccolith formation Ehud Bino , Lior Aram , Debojit Paul , Yuval Kadan , Daniel Clare , James B. Gilchrist , Nadav Elad , Assaf Gal Advanced Functional Materials 41 DOI: 10.1002/adfm.202415344 Diamond Proposal Number(s): [33808] Open Access Show Abstract ▼ Abstract: Inorganic minerals that form via regulated biological processes exhibit remarkable properties. This is due to the involvement of macromolecules that control biomineralization. Even though the interactions of these biopolymers with solid mineral phases are intensely studied, not much is known about their involvement in the preceding steps of intracellular transport of the mineral building blocks. In this work, the model system of coccolith calcite crystallization is utilized to address the role of mineral-associated polysaccharides in the transport of calcium ions. State-of-the-art cryo-electron tomography is used to image in situ ion-rich dense phases in the wild-type and in two mutant strains, defected in coccolith production. The results show that the abundance and solubility of the calcium-rich condensates need to be finely tuned for proper crystallization. When the native macromolecular assemblage is compromised, calcium is still present in the calcifying fluid as a solute, but this is not sufficient for coccolith development. These results suggest that biomineralizing systems achieve superior regulation of crystallization due to the use of dense macromolecule-rich phases.	Nov 2024
Krios IV-Titan Krios IV at Diamond Scios-Scios at Diamond	Intracellular morphogenesis of diatom silica is guided by local variations in membrane curvature Lior Aram , Diede De Haan , Neta Varsano , James B. Gilchrist , Christoph Heintze , Ron Rotkopf , Katya Rechav , Nadav Elad , Nils Kröger , Assaf Gal Nature Communications 15 DOI: 10.1038/s41467-024-52211-x Diamond Proposal Number(s): [29609] Open Access Show Abstract ▼ Abstract: Silica cell-wall formation in diatoms is a showcase for the ability of organisms to control inorganic mineralization. The process of silicification by these unicellular algae is tightly regulated within a membrane-bound organelle, the silica deposition vesicle (SDV). Two opposing scenarios were proposed to explain the tight regulation of this intracellular process: a template-mediated process that relies on preformed scaffolds, or a template-independent self-assembly process. The present work points to a third scenario, where the SDV membrane is a dynamic mold that shapes the forming silica. We use in-cell cryo-electron tomography to visualize the silicification process in situ, in its native-state, and with a nanometer-scale resolution. This reveals that the plasma membrane interacts with the SDV membrane via physical tethering at membrane contact sites, where the curvature of the tethered side of the SDV membrane mirrors the intricate silica topography. We propose that silica growth and morphogenesis result from the biophysical properties of the SDV and plasma membranes.	Sep 2024
I14-Hard X-ray Nanoprobe	Periplasmic bacterial biomineralization of copper sulfide nanoparticles Yeseul Park , Zohar Eyal , Péter Pekker , Daniel M. Chevrier , Christopher T. Lefèvre , Pascal Arnoux , Jean Armengaud , Caroline L. Monteil , Assaf Gal , Mihály Pósfai , Damien Faivre Advanced Science 18 DOI: 10.1002/advs.202203444 Diamond Proposal Number(s): [23693] Open Access Show Abstract ▼ Abstract: Metal sulfides are a common group of extracellular bacterial biominerals. However, only a few cases of intracellular biomineralization are reported in this group, mostly limited to greigite (Fe3S4) in magnetotactic bacteria. Here, a previously unknown periplasmic biomineralization of copper sulfide produced by the magnetotactic bacterium Desulfamplus magnetovallimortis strain BW-1, a species known to mineralize greigite (Fe3S4) and magnetite (Fe3O4) in the cytoplasm is reported. BW-1 produces hundreds of spherical nanoparticles, composed of 1–2 nm substructures of a poorly crystalline hexagonal copper sulfide structure that remains in a thermodynamically unstable state. The particles appear to be surrounded by an organic matrix as found from staining and electron microscopy inspection. Differential proteomics suggests that periplasmic proteins, such as a DegP-like protein and a heavy metal-binding protein, could be involved in this biomineralization process. The unexpected periplasmic formation of copper sulfide nanoparticles in BW-1 reveals previously unknown possibilities for intracellular biomineralization that involves intriguing biological control and holds promise for biological metal recovery in times of copper shortage.	Aug 2022

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