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Tips & tricks of note: Sample preparation: Using NMR analysis of point-mutants to choose an ideal cryoEM sample. Ideas for overcoming preferred orientation. Data processing: Understanding substrate specificity by solving multiple structures.    Alternative methods: Site-directed mutagenesis, NMR as a...

DNA polymerase ζ (Polζ) belongs to the same B-family as high-fidelity replicative polymerases, yet is specialized for the extension reaction in translesion DNA synthesis (TLS). Despite its importance in TLS, the structure of Polζ was unknown. We resolved structures of...

The sense of hearing and balance begins with the mechanosensory transduction (MT) channel, which converts mechanical stimuli into electrochemical signals, and which is typically localized within hair cells of the mammalian inner ear. Despite decades of effort focused on discovering...

Data processing: Practicalities of neural network picker software installation and usage. How to choose a particle picker & validation. Neural network picking for single particle analysis, filaments, & tomography. Interplay of particle picking and preferred orientations. Particle centering.

Genomic regions with high guanine content have the capacity to fold into four-stranded non-B DNA structures known as G-quadruplexes (G4s). Extensive bioinformatic inquiries have revealed that G4 motifs are conserved and non-randomly distributed throughout the genome. G4s are epigenetic features...

The success of any structural biology project is dependent on achieving sufficient purity and concentration of the sample-of-interest. Samples for cryo-EM generally have lower requirements for purity and concentration compared to other structural methods, making it a highly attractive approach...

LARGE1 is a bifunctional glycosyltransferase that polymerizes matriglycan – a linear polysaccharide of alternating xylose and glucuronic acid – on dystroglycan. Mutations in LARGE1 which decrease matriglycan can cause muscular dystrophies that are sometimes accompanied by intellectual disability. Matriglycan acts...

The ankyrin-1 complex tethers the spectrin-actin cytoskeleton to the red blood cell (RBC) membrane, and acts as a metabolic hub, connecting membrane proteins that are involved in gas exchange, pH control, and regulation of cellular volume and deformability. Mutations in...