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Many protein condensates can convert to fibrillar aggregates, but the underlying mechanisms are unclear. Liquid-liquid phase separation (LLPS) of spider silk proteins, spidroins, suggests a regulatory switch between both states. Here, we combine microscopy and native mass spectrometry to investigate the influence of protein sequence, ions, and regulatory domains on spidroin LLPS. We find that salting out-effects drive LLPS via low-affinity stickers in the repeat domains. Interestingly, conditions that enable LLPS simultaneously cause dissociation of the dimeric C-terminal domain (CTD), priming it for aggregation. Since the CTD enhances LLPS of spidroins but is also required for their conversion into amyloid-like fibers, we expand the stickers and spacers-model of phase separation with the concept of folded domains as conditional stickers that represent regulatory units.
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http://dx.doi.org/10.1021/acs.nanolett.3c00773 | DOI Listing |
J Biomol Struct Dyn
December 2024
Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi, India.
Nucleocapsid protein (N) of SARS-CoV-2 is a multivalent protein, which is responsible for viral replication, assembly, packaging and modulates host immune response. In this study, we report conformational measurements of N protein at different pH by observing transition in secondary and tertiary structural contents by biophysical and computational approaches. Spectroscopic measurements revealed that N protein loses its secondary and tertiary structure at extreme acidic pH while maintaining its native conformation at mild acidic and alkaline pH.
View Article and Find Full Text PDFEnviron Sci Process Impacts
December 2024
Department of Chemistry, University of Toronto, Canada.
Proton transfer reaction mass spectrometry (PTR-MS) is often employed to characterize gas-phase compounds in both indoor and outdoor environments. PTR-MS measurements are usually made without upstream chromatographic separation, so it can be challenging to differentiate between an ion of interest, its isomers, and fragmentation products from other species all detected at the same mass-to-charge ratio. These isomeric contributions and fragmentation interferences can confound the determination of accurate compound mixing ratios, the assignment of accurate chemical properties, and corresponding analyses of chemical fate.
View Article and Find Full Text PDFPhys Chem Chem Phys
December 2024
Semenov Federal Research Center for Chemical Physics, Kosygina, 4, 119991 Moscow, Russia.
The ability of particles to transform absorbed energy into translational movements brings peculiar order into nonequilibrium matter. Connected together into a chain, these particles collectively behave completely differently from well-known equilibrium polymers. Examples of such systems vary from nanoscale to macroscopic objects.
View Article and Find Full Text PDFSmall
December 2024
School of Materials and Energy, Yunnan University, Kunming, 650091, China.
Perovskite solar cells (PSCs) have achieved impressive efficiency, but their commercialization is limited by issues like chemical homogeneity within the perovskite films, leading to defects and phase segregation, which severely compromise the stability and performance of PSCs. This study presents a novel approach to overcoming these barriers by employing N,N-methylenebisacrylamide (MBA) as a multifunctional crosslinking agent within the perovskite structure. MBA enhances chemical uniformity both laterally and vertically, improves crystallinity, and boosts overall film stability by forming a robust crosslinked network that regulates nucleation and growth dynamics during the pre-seeding process.
View Article and Find Full Text PDFMol Cell Biol
December 2024
Department of Biology, University of Iowa, Iowa City, Iowa, USA.
Med15 is a general transcriptional regulator and tail module subunit within the RNA Pol II mediator complex. The Med15 protein has a well-structured N-terminal KIX domain, three activator binding domains (ABDs) and several naturally variable polyglutamine (poly-Q) tracts (Q1, Q2, Q3) embedded in an intrinsically disordered central region, and a C-terminal mediator association domain (MAD). We investigated how the presence of ABDs and changes in length and composition of poly-Q tracts influences Med15 activity using phenotypic, gene expression, transcription factor interaction and phase separation assays of truncation, deletion, and synthetic alleles.
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