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Roland L. Knorr

Publications and source records attributed to Roland L. Knorr.

4 recordsLinked to original sources

Condensate-mediated shape transformations of cellular membranes by capillary forces

Phase-separated biomolecular condensates with liquid-like properties play a key role in the organization and compartmentalization of the intracellular environment. Condensate-mediated capillary forces acting on membranes drive physiologically important reshaping of membrane-bound organelles, such as vacuoles and autophagosomes. Here, we explore condensate-mediated membrane shape transformations. We employ {\textit{in planta}} live-cell imaging, an \textit{in vitro} reconstitution system with tunable interfacial tension, and computer simulations of an elastic membrane model to describe three morphologies of membrane structures localized at condensate interfaces: tubes, sheets, and cups. We find that the forces associated with high interfacial tension drive the formation of stable sheets, while tubes and cups prevail at lower interfacial tension. We calculate the free energies of each membrane shape and identify the energy barriers that govern the transitions between the shapes. With this approach, we find that shape transformations depend on the history of the interfacial membrane and exhibit a tube-to-cup hysteresis. These findings indicate that temporal control of condensate surface properties can mediate the morphogenesis of cup-like structures in cells, such as the formation of "bulbs" within plant vacuoles. Our results further generalize how the interplay of condensates and membranes contributes to intracellular organization.

cond-mat.soft↗

Modeling membrane morphological change during autophagosome formation

Autophagy is an intracellular degradation process that is mediated by de novo formation of autophagosomes. Autophagosome formation involves dynamic morphological changes; a disk-shaped membrane cisterna grows, bends to become a cup-shaped structure, and finally develops into a spherical autophagosome. We have constructed a theoretical model that integrates the membrane morphological change and entropic partitioning of putative curvature generators, which we have used to investigate the autophagosome formation process quantitatively. We show that the membrane curvature and the distribution of the curvature generators stabilize disk- and cup-shaped intermediate structures during autophagosome formation, which is quantitatively consistent with in vivo observations. These results suggest that various autophagy proteins with membrane curvature-sensing properties control morphological change by stabilizing these intermediate structures. Our model provides a framework for understanding autophagosome formation.

physics.bio-ph↗

Micron-sized domains in quasi single-component giant vesicles

Giant unilamellar vesicles (GUVs), are a convenient tool to study membrane-bound processes using optical microscopy. An increasing number of studies highlights the potential of these model membranes when addressing questions in membrane biophysics and cell biology. Among them, phase transitions and domain formation, dynamics and stability in raft-like mixtures are probably some of the most intensively investigated. In doing so, many research teams rely on standard protocols for GUV preparation and handling involving the use of sugar solutions. Here, we demonstrate that following such a standard approach can lead to abnormal formation of micron-sized domains in GUVs grown from only a single phospholipid. The membrane heterogeneity is visualized by means of a small fraction (0.1 mol%) of a fluorescent lipid dye. For dipalmitoylphosphatidylcholine GUVs, different types of membrane heterogeneities were detected. First, an unexpected formation of micron-sized dye-depleted domains was observed upon cooling. These domains nucleated about 10 K above the lipid main phase transition temperature, TM. In addition, upon further cooling of the GUVs down to the immediate vicinity of TM, stripe-like dye-enriched structures around the domains are detected. The micron-sized domains in quasi single-component GUVs were observed also when using two other lipids. Whereas the stripe structures are related to the phase transition of the lipid, the dye-excluding domains seem to be caused by traces of impurities present in the glucose. Supplementing glucose solutions with nm-sized liposomes at millimolar lipid concentration suppresses the formation of the micron-sized domains, presumably by providing competitive binding of the impurities to the liposome membrane in excess. It is likely that such traces of impurities can significantly alter lipid phase diagrams and cause differences among reported ones.

q-bio.BM↗

Formation of autophagosomes coincides with relaxation of membrane curvature

Autophagy is an intracellular degradation process that employs complex membrane dynamics to isolate and break down cellular components. However, many unanswered questions remain concerning remodeling of autophagic membranes. Here, we focus on the advantages of theoretical modelling to study the formation of autophagosomes and to understand the origin of autophagosomal membranes. Starting from the well-defined geometry of final autophagosomes we ask the question of how these organelles can be formed by combining various preautophagosomal membranes such as vesicles, membrane tubules or sheets. We analyse the geometric constraints of autophagosome formation by taking the area of the precursor membranes and their internal volume into account. Our results suggest that vesicle fusion contributes little to the formation of autophagosomes. In the second part we quantify the curvature of the precursors and report that the formation of autophagosomes is associated with a strong relaxation of membrane curvature energy. This effect we find for a wide range of membrane asymmetries. It is especially strong for small distances between both autophagosomal membranes, as observed in vivo. We quantify the membrane bending energies of all precursors by considering membrane asymmetries. We propose that the generation and supply of pre-autophagosomal membranes is one limiting step for autophagosome formation.

q-bio.SC↗