Proteomics was employed to elucidate the intricate molecular mechanisms underlying the stress response of N. thermophilus when confronted with the synergistic challenges of high salt, alkaline pH, and elevated temperatures (HSAT conditions). This pioneering study presents the first comprehensive protein-level blueprint for polyextremophiles, uncovering a sophisticated interplay and synergy among halophilic, alkaliphilic, and thermophilic adaptive strategies. Our discoveries not only enrich the understanding of extremophile biology but also open up exciting avenues for exploring the resilience of life in extraterrestrial ecosystems, thereby offering invaluable prospects for astrobiological research.