bioRxiv Science⌕ Search

Biology subjects

Cioci, G.

Publications and source records attributed to Cioci, G..

3 recordsLinked to original sources

A Generative Neuro-Symbolic AI for Protein Sequence Design

Deep learning has revolutionized computational protein design, enabling the generation of sequences that fold onto target backbones with unprecedented accuracy. However, state-of-the-art inverse folding tools largely rely on auto-regressive sampling. While powerful, this paradigm is increasingly recognized for its inability to "think ahead", a crucial capacity to reliably create the complex, long-range inter-residue dependencies essential for most biological functions. To overcome these fundamental limitations, we introduce EffieDes, a generative neuro-symbolic AI framework that synergizes the predictive capabilities of deep learning with the logical precision of automated reasoning. EffieDes leverages deep learning to encode the target backbones fitness landscape into Effie-- a fully decomposable probabilistic graphical model (Potts model). This landscape is then rigorously explored by an automated reasoning prover to identify sequences that simultaneously satisfy complex design constraints and optimize backbone fitness. We validated this neuro-symbolic approach through the design of orthogonal sequence pairs that adopt identical folds but exhibit selective self-assembly, as well as the design of a de novo selective nanobody with nanomolar affinity for an immune-evasive SARS-CoV-2 variant. EffieDes provides a robust architecture for precisely dissecting learned fitness landscapes, offering a new path toward proteins with highly optimized performances and sophisticated functional objectives.

bioengineering↗

Structural investigations of the glucan water dikinase 1 mechanism and flexibility

Glucan-water-dikinase 1 (GWD1) plays an essential role in regulating starch metabolism in plants via O-6 phosphorylation of amylopectin. Here, we used biochemical characterization, AlphaFold2 modeling, X-ray crystallography and Small-Angle X-ray Scattering (SAXS) experiments to study its structure and catalytic mechanism. The protein is organized into five domains with two carbohydrate-binding modules (CBMs) at its N-terminal end followed by a central domain, whose structure was solved by X-ray crystallography in open and closed conformations. Next comes the domain carrying the catalytic histidine and the ATP-binding domain. We studied the spatial arrangement of the full enzyme and of several truncated forms by SAXS-driven modeling and identified a pivoting movement of the Histidine domain consistent with the enzymes autophosphorylation and subsequent phosphate transfer to a glucan. Our data suggest important residues at the domain interfaces that might assist catalysis and we hypothesize that the second CBM helps maintaining the catalytic domain close to the glucan chain for productive phosphate transfer. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/704335v1_ufig1.gif" ALT="Figure 1"> View larger version (46K): org.highwire.dtl.DTLVardef@1b860e5org.highwire.dtl.DTLVardef@1e172dcorg.highwire.dtl.DTLVardef@3c03edorg.highwire.dtl.DTLVardef@25c0d4_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

Designing symmetrical multi-component proteins using a hybrid generative AI approach

Proteins, the fundamental building blocks of biological function, orchestrate complex cellular processes by assembling into intricate structures through meticulous interactions. The design of specific protein-protein interfaces to create customized protein assemblies holds immense potential for various biotechnological applications. To address the current limitations in designing heteromeric interactions for multi-component assemblies, we developed a hybrid generative AI design approach. This method combines deep learning and automated reasoning, explicitly considering both positive and negative interaction states to favor heteromeric desired over undesired interactions. The approach leverages Effie, a deep-learned pairwise decomposable scoring function, and an advanced reasoning tool extended for multicriteria optimization of this function. Here, we tested the ability of this hybrid AI method to redesign homomeric interfaces of bacterial microcompartment components (BMC-H) into heteromeric assemblies. We benchmarked its performance against ProteinMPNN, a sequence design autoregressive model. Our in silico assessment, complemented by experimental validation, highlights the outperformance of the hybrid AI generative design approach, and its potential to unlock the engineering of complex multi-component self-assembling protein entities.

bioengineering↗