Abstract In spite of their great importance in biology, methods providing access to spontaneous molecular interactions with and on biological membranes have been sparse. The recent advent mass photometry quantify distributions unlabeled biomolecules landing surfaces raised hopes that this approach could be transferred membranes. Here, by introducing a new interferometric scattering (iSCAT) image processing analysis strategy adapted diffusing particles, we enable mass-sensitive particle...

Cell division is spatiotemporally precisely regulated, but the underlying mechanisms are incompletely understood. In social bacterium Myxococcus xanthus, PomX/PomY/PomZ proteins form a single megadalton-sized complex that directly positions and stimulates cytokinetic ring formation by tubulin homolog FtsZ. Here, we study structure mechanism of this in vitro vivo. We demonstrate PomY forms liquid-like biomolecular condensates phase separation, while PomX self-assembles into filaments...

Patterns formed by reaction-diffusion mechanisms are crucial for the development or sustenance of most organisms in nature. include dynamic waves, but more often found as static distributions, such animal skin patterns. Yet, a simplistic biological model system to reproduce and quantitatively investigate patterns has been missing so far. Here, we demonstrate that Escherichia coli Min system, known its oscillatory behavior between cell poles, is under certain conditions capable transitioning...

Modules that switch protein-protein interactions on and off are essential to develop synthetic biology; for example, construct orthogonal signaling pathways, control artificial protein structures dynamically, localization in cells or protocells. In nature, the E. coli MinCDE system couples nucleotide-dependent switching of MinD dimerization membrane targeting trigger spatiotemporal pattern formation. Here we present a de novo peptide-based molecular toggles reversibly between monomer dimer...

Single-particle plasmon spectroscopy has become a standard technique to detect and quantify the presence of unlabeled macromolecules. Here, we extend this method determine their exact distance from sensors with sub-nanometer resolution by systematically varying sensing range into surrounding adjusting size plasmonic nanoparticles. We improved current single-particle record continuously for hours scattering spectra thousands nanoparticles different sizes simultaneously 1.8 s time resolution....

The formation of large-scale patterns through molecular self-organization is a basic principle life. Accordingly, the engineering protein and gradients prime relevance for synthetic biology. As paradigm such pattern formation, bacterial MinDE system based on ATPase MinD ATPase-activating MinE lipid membranes. Min can be tightly regulated by tuning physical or biochemical parameters. Among biochemically engineerable modules, MinD's membrane targeting sequence, despite being key regulating...

Abstract Neurodevelopmental disorders are heterogeneous and identifying shared genetic aetiologies converging signalling pathways affected could improve disease diagnosis treatment. Truncating mutations of the abnormal spindle-like microcephaly associated ( ASPM ) gene cause autosomal recessive primary (MCPH) in humans. is a positive regulator Wnt/β-Catenin controls symmetric to asymmetric cell division. This process balances neural progenitor proliferation with differentiation during...

Abstract As one of the key elements in bacterial cell division, cytoskeletal protein FtsZ appears to be highly involved circumferential treadmilling along inner membrane, yielding circular vortices when transferred flat membranes. However, it remains unclear how a membrane-targeted can produce these dynamics. Here, we dissect roles membrane binding, GTPase activity, and unstructured C-terminal linker on chimera through vitro reconstitution different FtsZ-YFP-mts variants supported In...

Reversible membrane targeting of proteins is one the key regulators cellular interaction networks, for example, signaling and polarization. So-called "membrane switches" are thus highly attractive targets design minimal cells but have so far been tricky to reconstitute in vitro. Here, we introduce cell-free prenylated protein synthesis (CFpPS), which enables a single reaction mix including prenylation machinery. CFpPS can confer affinity any via addition 4-peptide motif its C-terminus offers...

Abstract BACKGROUND Chemically defined media (CDM) are used routinely in industrial settings for the production of biopharmaceuticals from mammalian cells, so attention has shifted to understanding their chemical behavior and impact on process robustness. In this context, one particular problem is risk cross reactivity instability complex mixtures chemicals at high concentrations. RESULTS order characterize potentially unstable medium compounds feed media, we developed validated a novel...

Summary Cell division is spatiotemporally precisely regulated, but the underlying mechanisms are incompletely understood. In social, predatory bacterium Myxococcus xanthus , PomX/PomY/PomZ proteins form a single large megadalton-sized complex that directly positions and stimulates cytokinetic ring formation by tubulin homolog FtsZ. Here, we studied structure mechanism of this in vitro vivo. We demonstrate PomY forms liquid-like biomolecular condensates phase separation, while PomX...

Abstract A better alignment of preclinical and clinical neurobiological measures could help improve neuropsychiatric disease therapeutic development. This unit describes a compendium hypothesis‐driven neuroanatomical phenotyping strategies to be employed in genetic mouse models. Using disease‐based alterations as guide, these are histological immunohistochemical methodologies also applied human tissue. They include quantification assays neurochemical‐, newly born neuron‐ glial‐cell markers,...

Short-lived or transient interactions of macromolecules at and with lipid membranes, an interface where a multitude essential biological reactions take place, are inherently difficult to assess standard biophysical methods. The introduction mass-sensitive particle tracking (MSPT) constitutes important step toward thorough quantitative characterization such processes. Technically, this was made possible through the advent interferometric scattering microscopy (iSCAT)-based mass photometry...

Abstract In spite of their great importance in biology, methods providing access to spontaneous molecular interactions with and on biological membranes have been sparse. So far, it has consensus that observation sufficient sensitivity time resolution requires the introduction - predominantly fluorescent-labels system. However, recent advent mass photometry quantify distributions unlabelled biomolecules landing surfaces raised hopes this approach could be transferred membranes. Here, we...

As a key mechanism underpinning many biological processes, protein self-organization has been extensively studied. However, the potential to apply distinctive, nonlinear biochemical properties of such self-organizing systems biotechnological problems as facile detection and characterization biomolecular interactions not yet explored. Here, we describe an in vitro assay 96-well plate format that harnesses emergent behavior Escherichia coli Min system provide readout interactions. Crucial for...

Short-lived or transient interactions of macromolecules at and with lipid membranes, an interface where a multitude essential biological reactions take place, are inherently difficult to assess standard biophysical methods. The introduction mass-sensitive particle tracking (MSPT) constitutes important step toward thorough quantitative characterization such processes. Technically, this was made possible through the advent interferometric scattering microscopy (iSCAT)-based mass photometry...

Abstract Bottom-up synthetic biology is a powerful tool for uncovering the mechanisms underlying vital biological processes, such as signaling and cell polarization. The core principle of reconstituting cellular functions in their minimal forms can be achieved through modular protein design. However, assembling multiple purified proteins into functional synchronized system remains technical challenge. fact that many regulatory show direct or indirect membrane interactions further exacerbates...

ABSTRACT Short-lived or transient interactions of macromolecules at and with lipid membranes, an interface where a multitude essential biological reactions take place, are inherently difficult to assess standard biophysical methods. The introduction mass-sensitive particle tracking (MSPT) constitutes important step towards thorough quantitative characterization such processes. Technically, this was made possible through the advent interferometric scattering microscopy (iSCAT)-based mass...