Not a chocolate easter egg, but rather Xenopus egg extract (XRCC4-like factor, XLF) on SEC-MALS!
Comments: Happy Easter and Passover! Here is a fun paper using @Sepax SRT SEC-300 Light Scattering for #HPLC SEC-MALS analysis of Xenopus egg extract (XRCC4-like factor, XLF) published by @Harvard Medical School. Thank you to the authors and supporting scientists!
Analytical SEC HPLC of NHEJ1 with Sepax SRT SEC-300, Light Scattering
XLF acts as a flexible connector during non-homologous end joining
Company Name:?Harvard Medical School
Year:?2020
Product:?HPLC Column, Sepax, SRT SEC-300, 5 um, 7.8x300mm, Light Scattering SEC
Part Number:?215300LS-7830
Separation Mode:?Size Exclusion Chromatography (SEC) coupled with Multi Angle Light Scattering (MALS)
Broad Sample Category:?Protein
领英推荐
Specific Sample:?XLF C-terminal tail in NHEJ synaptic complex assembly
Mobile Phase:?50 mM Na-MES, pH 6.5, 350 mM Tris, 10% glycerol, and 1 mM DTT
Instrument:?HPLC; SEC-MALS
Full Title:?XLF acts as a flexible connector during non-homologous end joining
Citation:?Carney, Sean M., et al. "XLF acts as a flexible connector during non-homologous end joining." Elife 9 (2020): e61920. https://doi.org/10.7554/eLife.61920
Abstract:?Non-homologous end joining (NHEJ) is the predominant pathway that repairs DNA double-strand breaks in vertebrates. During NHEJ DNA ends are held together by a multi-protein synaptic complex until they are ligated. Here, we use?Xenopus laevis?egg extract to investigate the role of the intrinsically disordered C-terminal tail of the XRCC4-like factor (XLF), a critical factor in end synapsis. We demonstrate that the XLF tail along with the Ku-binding motif (KBM) at the extreme C-terminus are required for end joining. Although the underlying sequence of the tail can be varied, a minimal tail length is required for NHEJ. Single-molecule FRET experiments that observe end synapsis in real-time show that this defect is due to a failure to closely align DNA ends. Our data supports a model in which a single C-terminal tail tethers XLF to Ku, while allowing XLF to form interactions with XRCC4 that enable synaptic complex formation.
Link to Paper:?https://elifesciences.org/articles/61920
Thank you?to the authors and supporting scientists!