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Biology subjects

Hardy, J. M.

Publications and source records attributed to Hardy, J. M..

2 recordsLinked to original sources

ProteinDJ: a high-performance and modular protein design pipeline

Leveraging artificial intelligence and deep learning to generate proteins de novo (a.k.a. synthetic proteins) has unlocked new frontiers of protein design. Deep learning models trained on protein structures can generate novel protein designs that explore structural landscapes unseen by evolution. This approach enables the development of bespoke binders that target specific proteins and domains through new protein-protein interactions. However, successful binder generation can suffer from low in silico success rates, often requiring thousands of designs and hundreds of GPU hours to obtain enough hits for experimental testing. While workstation implementations are available for binder design, these are limited in both scalability and throughput. There is a lack of efficient open-source protein design pipelines for high-performance computing (HPC) systems that can maximise hardware resources and parallelise the workflow efficiently. Here, we present ProteinDJ--an implementation of a synthetic protein design workflow that is deployable on HPC systems using the Nextflow portable workflow management system and Apptainer containerisation. It parallelises the workload across both GPUs and CPUs, facilitating generation and testing of hundreds of designs per hour, accelerating the discovery process. ProteinDJ is designed to be modular and includes RoseTTAFold Diffusion (RFdiffusion) or BindCraft for fold generation, ProteinMPNN or Full-Atom MPNN (FAMPNN) for sequence design, and AlphaFold2 or Boltz-2 for prediction and validation of designs and binder-target interfaces, with supporting packages for structural evaluation of designs. ProteinDJ democratises protein binder design through its robust and user-friendly implementation and provides a framework for future protein design pipelines. ProteinDJ is freely available at https://github.com/PapenfussLab/proteindj.

bioinformatics↗

When two is a crowd - Structural mapping of PEAK pseudokinase interactions identifies 14 3 3 as a molecular switch for PEAK3/Crk signaling.

PEAK pseudokinases regulate cell migration, invasion and proliferation by recruiting key signaling proteins to the cytoskeleton. Despite lacking catalytic activity, alteration in their expression level is associated with several aggressive cancers. Here, we elucidate new molecular details of key PEAK signaling interactions with the adapter proteins CrkII and Grb2 and the scaffold protein 14-3-3. Our findings rationalize why the dimerization of PEAK proteins has a crucial function in signal transduction and provide biophysical and structural data to unravel binding specificity within the PEAK interactome. We identify a conserved high affinity 14-3-3 motif on PEAK3 and demonstrate its role as a molecular switch to regulate CrkII binding. Together, our studies provide a detailed structural snapshot of PEAK interaction networks and further elucidate how PEAK proteins, especially PEAK3, act as dynamic scaffolds that exploit adapter proteins to control signal transduction in cell growth/motility and cancer.

biochemistry↗