Abstract Allosteric modulation enables precise control of protein activity but remains difficult to harness for selective inhibitor design. Traditional high‐throughput screening for allosteric modulators is still costly and time‐consuming, underscoring the need for predictive computational approaches. Here, we combined network and shortest‐path analyses to predict interprotomer communication nodes that regulate the pro‐apoptotic activity of human galectin‐7 (GAL‐7). We identify a minimal electrostatic network (R20‐R22‐D103) as a key allosteric node controlling dimer stability and signal transmission between the two distant glycan binding sites. Our predictions guided the engineering of four variants (R20A, R22A, D103A, and R20A‐R22A), all of which impaired GAL‐7‐induced apoptosis in human T cells. Biophysical and structural analyses confirmed that disrupting the R20‐D103 interaction weakens interprotomer communication and destabilizes the dimer, while compensatory edges partially restore connectivity. These results demonstrate that residue‐network fingerprinting enables predictive mapping of global communication pathways and reveal R20, R22, and D103 as key allosteric determinants of GAL‐7 function. The integrative framework introduced here can be extended to identify and exploit allosteric communication pathways in other homodimeric proteins, offering a generalizable strategy for rational modulator design.
Pham et al. (Tue,) studied this question.