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Synthetic nanobodies-also called sybodies-have proven valuable for stabilizing conformations of purified proteins, advancing structural and functional studies for example of transmembrane proteins. However, their utility in modulating protein function in living cells has remained less well explored. Structural Maintenance of Chromosomes (SMC) complexes facilitate chromosome organization, a fundamental process in all domains of life. In this study, we target the bacterial SMC complex, Smc-ScpAB, in Bacillus subtilis with synthetic nanobodies, aiming to identify key functional regions of the protein complex in a largely unbiased manner. We first isolate sybodies that specifically bind purified Smc-ScpAB and then express them in B. subtilis to select binders capable of disrupting Smc-ScpAB function, leading to chromosome segregation defects and cell death. Mapping and biochemical characterization show that the 14 disruptive sybodies belong to one of three library designs, target the Smc subunit near the same coiled coil arm interface and modulate its ATPase activity in two principal ways, highlighting the mid-region of the Smc coiled coil as critical feature of the SMC-DNA folding process. These findings underscore the potential of sybodies-and, by extension, designed binders-as versatile tools for probing dynamic protein function in living cells.
Pubmed ID: 42153619
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Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
View all literature mentionsIndependent body that funds investigator-driven frontier research in the European Union (EU). It is part of the Seventh Research Framework Programme (FP7).
View all literature mentionsThe Swiss National Science Foundation (SNSF) is Switzerland''s leading provider of scientific research funding. The SNSF annually supports some 7200 researchers, almost 80 percent of whom are aged 35 years or younger. With its federal mandate, it supports basic research in all disciplines, from philosophy and biology to the nanosciences and medicine. It also invests in applied research in various scientific fields. The focus of its activities is the scientific endorsement of projects submitted by researchers. The best applicants are funded by the SNSF with an annual total amount equalling approximately CHF 700 million. Established in 1952 as a foundation under private law, the SNSF has the autonomy it needs to promote independent scientific research. The SNSF is committed to promoting young scientists and works to ensure that scientific research in Switzerland has the most favourable conditions for developing internationally. It also encourages dialogue between scientists and representatives in society, politics and the economy.
View all literature mentionsThis monoclonal targets c-Myc
View all literature mentionsStatistical analysis software that combines scientific graphing, comprehensive curve fitting (nonlinear regression), understandable statistics, and data organization. Designed for biological research applications in pharmacology, physiology, and other biological fields for data analysis, hypothesis testing, and modeling.
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