Protein complexes are key players in development, functioning as molecular machines whose proper assembly is crucial for numerous cellular processes. Their formation at the correct time and location is facilitated by specialized scaffold proteins. A prominent example is the mechanosensitive cytoskeletal protein zyxin. Zyxin is of particular interest due to its role in assembling actin filaments within focal adhesion complexes and transducing signals from the cytoskeleton to the genetic apparatus. In this study, we employed co-immunoprecipitation coupled with mass spectrometry to analyze changes in zyxin-associated protein complexes from lysates of Xenopus frog embryos across three early developmental stages: early gastrulation, early neurulation, and late neural fold stages. Our analysis revealed stage-specific alterations in the isoform repertoires of several established zyxin partners, including components of the focal adhesion complex, transcriptional regulators, and proteins involved in its phosphorylation. We also identified changes in the phosphorylation patterns of different zyxin isoforms. Collectively, these findings provide a valuable resource for elucidating the function of the scaffold protein zyxin and its role in embryonic tissue development. Furthermore, the identification of kinases and caspases within the zyxin interactome suggests its involvement in key signaling pathways related to cell survival and apoptosis. Finally, confirmed interactions with partners such as 14-3-3 proteins, Yap, and TEF-1 highlight the multifaceted role of zyxin in cellular regulation and its potential as a therapeutic target.