Verification: model-free phasing with enhanced predicted models in ARCIMBOLDO_SHREDDER

The accuracy of the models predicted using AlphaFold and RoseTTAFold eases phasing with molecular replacement and raises the question of model bias. ARCIMBOLDO_SHREDDER solves structures while aiming to establish the experimental information in a crystallographic determination by introducing systematic, local verification of phasing solutions.

Exiting the tunnel of uncertainty: crystal soak to validated hit

A brief overview of techniques used in the hit-identification/validation stage of drug discovery is given, highlighting the importance of orthogonal measurements and triangulation in early-stage crystallographic hit validation.

Putting AlphaFold models to work with phenix.process_predicted_model and ISOLDE

phenix.process_predicted_model and ISOLDE provide seamless integration of AlphaFold-predicted models into protein structure-solution workflows.

A round-robin approach provides a detailed assessment of biomolecular small-angle scattering data reproducibility and yields consensus curves for benchmarking

Small-angle X-ray scattering (SAXS) and small-angle neutron scattering (SANS) measurements of five standard proteins in solution using 12 SAXS and four SANS instruments demonstrate reproducibility and yield consensus scattering profiles that provide a foundation benchmarking set to evaluate approaches to scattering-profile prediction from atomic coordinates.

Polyelectrolyte coating of cryo-EM grids improves lateral distribution and prevents aggregation of macromolecules

Coating cryo-EM grids with single-stranded DNA before applying the sample reduces the aggregation of macromolecules and improves their distribution. It enables the determination of structures from samples that aggregate on conventional grids.

Zymogenic latency in an ∼250-million-year-old astacin metallopeptidase

The horseshoe crab Limulus polyphemus is an ancient chelicerate that is a model organism for the study of the evolution and function of peptidases. It contains a member of the astacin metallopeptidases, the mechanism of latency of which was revealed by X-ray structural analysis.

Glycoside hydrolase subfamily GH5_57 features a highly redesigned catalytic interface to process complex hetero-β-mannans

A phylogenetically distant glycoside hydrolase family 5 member retrieved from the capybara gut microbiome displays a large and open catalytic interface that is adapted to recognize complex heteromannans, establishing the novel GH5_57 subfamily. This study expands the current mechanistic and functional understanding of one of the largest GH families.

Structural and functional investigation of the human snRNP assembly factor AAR2 in complex with the RNase H-like domain of PRPF8

The crystal structure of human AAR2 bound to the central spliceosomal factor PRPF8 and in vitro functional data yield insights into the structural basis of snRNP assembly in humans.

Crystal structure of DNA polymerase I from Thermus phage G20c

The crystal structure of DNA polymerase I from Thermus phage G20c is presented and represents the first crystal structure of DNA polymerase I from a group of related thermophilic bacteriophages. The structure reveals a new structural motif termed SβαR that was not previously identified in other DNA polymerases I (or in the DNA polymerase A family).

Unravelling the regulation pathway of photosynthetic AB-GAPDH

Regulation of the heteromeric form of photosynthetic glyceraldehyde 3-phosphate dehydrogenase (AB-GAPDH) depends on oscillation between a fully active heterotetramer (A₂B₂) and inhibited oligomers. Experimental evidence demonstrates that the inhibition of spinach AB-GAPDH depends on the formation of dimers, tetramers or pentamers of A₂B₂ modules linked together by C-terminal extensions of the B subunits that extend from one modular tetramer and occupy two active sites of the adjacent tetramer.