Fine-Tuning Protein Language Models on Human Spatial Constraint Yields State-of-the-Art Variant Effect Prediction

Protein language models (PLMs) achieve state-of-the-art performance in predicting effects of missense variants, yet they do not explicitly consider variation within the human population. Here, we introduce Human Spatial Constraint (HuSC), a framework for quantifying intraspecies constraint on missense variants that integrates population-scale human genetic variation with 3D protein structures. We then fine-tune PLMs on HuSC scores. HuSC models the expected frequency of missense variation under neutral evolution and compares it to observed variation, accounting for both variation in mutational processes and 3D structural context. HuSC outperforms traditional inter- and intraspecies conservation metrics in predicting pathogenic variants. By focusing on intraspecies variation, HuSC reveals protein sites under human-specific constraint that cannot be captured by interspecies models. Integrating this intraspecies perspective into PLMs by fine-tuning on HuSC scores improves the prediction of variant fitness from deep mutational scans across diverse taxa and functional assay types. The improvement after fine-tuning comes largely from reducing bias toward wild-type sequences in regions that tolerate variation. Together, these results demonstrate that combining intraspecies constraint with cross-species PLMs improves their performance in variant-effect interpretation.