Par3与OGDH直接相互作用验证：如何评估原生条件下蛋白质结合证据？

The study does not demonstrate direct binding between full-length Par3 and full-length OGDH under native conditions. While fragment-based GST pull-downs (Figure 2K) and interface mutagenesis (Figures 2L–2O) argue for direct contact, co-immunoprecipitations (Figures 2E–2F) and PLA (Figure 2D) cannot exclude bridging proteins, and fragments may engage non-physiological surfaces. The literature reveals no prior characterization of PARD3-OGDH interactions [1][2], and no studies have applied rigorous biophysical validation methods for similar polarity protein-metabolic enzyme interactions. Without biophysical binding data using full-length proteins or crosslinking in intact mitochondria, a direct, native interaction remains inferred rather than demonstrated.