MOTIVATION: RNA-protein interactions play essential roles in every living organism, with RNA transcription, processing, and translation being just a few examples. Therefore, determining the set of RNAs that are bound by individual RNA-binding proteins, as well as the precise location of the interaction, is crucial for biological understanding. CLIP (UV-cross-linking and immunoprecipitation) is a method developed to study these interactions. Several variations of the CLIP protocol have been developed, e.g. iCLIP (individual-nucleotide resolution CLIP), which offers nucleotide-precise resolution of the cross-linking event. RESULTS: PARANOiD is a versatile software for fully automated analysis of iCLIP and iCLIP2 data. It contains all steps necessary for preprocessing, the determination of cross-link locations, and several additional steps, which can be used to detect specific characteristics, e.g. definite distances between cross-link events or identify binding motifs. Additionally, results are visualized as statistical plots for a quick overview and as standardized bioinformatics file formats, which can be used for further analysis steps. AVAILABILITY AND IMPLEMENTATION: PARANOiD is published under the MIT license and is available from https://github.com/patrick-barth/PARANOiD. The documentation is available at https://paranoid.readthedocs.io/en/latest/index.html.

Generating diverse, all‐atom conformational ensembles of dynamic proteins such as G‐protein‐coupled receptors (GPCRs) is critical for understanding their function, yet most generative models simplify atomic detail or ignore conformational diversity altogether. We present latent diffusion for full protein generation (LD-FPG), a framework that constructs complete all‐atom protein structures, including every side‐chain heavy atom, directly from molecular dynamics (MD) trajectories. LD-FPG employs a Chebyshev graph neural network (ChebNet) to obtain low‐dimensional latent embeddings of protein conformations, which are processed using three pooling strategies: blind, sequential and residue‐based. A diffusion model trained on these latent representations generates new samples that a decoder, optionally regularized by dihedral‐angle losses, maps back to Cartesian coordinates. Using D2R-MD, a $2\mu\text{s}$ MD trajectory (12 000 frames) of the human dopamine D$2$ receptor in a membrane environment, the sequential and residue-based pooling strategies reproduce the reference ensemble with high structural fidelity (all‐atom lDDT \~ $0.7$; $C\alpha$-lDDT \~ $0.8$) and recovers backbone and side‐chain dihedral‐angle distributions with a Jensen–Shannon divergence $<0.03$ compared to the MD data. LD-FPG thereby offers a practical route to system‐specific, all‐atom ensemble generation for large proteins, providing a promising tool for structure‐based therapeutic design on complex, dynamic targets. The D2R-MD dataset and our implementation are freely available to facilitate further research.

Eukaryotic transmembrane helical (TMH) proteins perform a wide diversity of critical cellular functions, but remain structurally largely uncharacterized and their high-resolution structure prediction is currently hindered by the lack of close structural homologues. To address this problem, we present a novel and generic method for accurately modeling large TMH protein structures from distant homologues exhibiting distinct loop and TMH conformations. Models of the adenosine A2AR and chemokine CXCR4 receptors were first ranked in GPCR-DOCK blind prediction contests in the receptor structure accuracy category. In a benchmark of 50 TMH protein homolog pairs of diverse topology (from 5 to 12 TMHs), size (from 183 to 420 residues) and sequence identity (from 15% to 70%), the method improves most starting templates, and achieves near-atomic accuracy prediction of membrane-embedded regions. Unlike starting templates, the models are of suitable quality for computer-based protein engineering: redesigned models and redesigned X-ray structures exhibit very similar native interactions. The method should prove useful for the atom-level modeling and design of a large fraction of structurally uncharacterized TMH proteins from a wide range of structural homologues.