In this study, collagen hydrolysate was prepared from chicken skin which is one of the most underutilized poultry by-products. Its cryoprotective effect against protein freeze-denaturation was evaluated on the natural actin-myosin complex (actomyosin, NAM) obtained from chicken breast muscle. This comprehensive study was aimed to elucidate the mechanism of chicken collagen hydrolysate (CCH) in retarding protein freeze-denaturation, starting from the inhibitive effect of CCH on ice growth, followed by analyzing its cryoprotective action on NAM conformation and oxidation after frozen storage. Finally, the influence of these modifications on protein gel formation and in vitro digestibility was also investigated as functional tests relevant to the food industry.