Molecular biology aims to understand the molecular basis of cellular responses, unravel dynamic regulatory networks, and model complex biological systems. However, these studies remain challenging in non-model species as a result of poor functional annotation of regulatory proteins, like kinases or phosphatases. To overcome this limitation, we developed a multi-layer neural network that annotates proteins by determining functionality directly from the protein sequence. We annotated the kinases and phosphatases in the non-model species, Glycine max (soybean), achieving a prediction sensitivity of up to 97%. To demonstrate the applicability, we used our functional annotations in combination with Bayesian network principles to predict signaling cascades using time series phosphoproteomics. We shed light on phosphorylation cascades in soybean seedlings upon cold treatment and identified Glyma.10G173000 (TOI5) and Glyma.19G007300 (TOT3) as key temperature response regulators in soybean. Importantly, the signaling cascade predictions do not rely upon known upstream kinases, kinase motifs, or protein interaction data, enabling de novo identification of kinase-substrate interactions. In addition to high accuracy and strong generalization, we showed that our functional prediction neural network is scalable to other model and non-model species, including Oryza sativa (rice), Zea mays (maize), Sorghum bicolor (sorghum), and Triticum aestivum (wheat). Overall, we demonstrated a data-driven systems biology approach for non-model species leveraging our predicted upstream kinases and phosphatases.