Supplement Summary I am requesting an administrative supplement for the purchase of a Cytiva ÄKTA Pure fast protein liquid chromatography (FPLC) system. This FPLC system is capable of operating a variety of chromatography columns, including preparative and analytical size exclusion, ion exchange and affinity columns. This equipment will be crucial for completing the goals of my NIGMS R35 grant (R35GM144341) by allowing us to conduct detailed biochemical and hydrodynamic analyses of at least three key protein complexes involved in the Wnt5a- Ror morphogenetic pathway. Specifically, 1) during the course of our study of WNT5A mutations found in Robinow syndrome patients, we discovered that the mutant proteins form aberrant disulfide bonds that result in WNT5a dimerization or multimerization. High-resolution size exclusion chromatography is required to further analyze the formation of these higher-order Wnt5a oligomers, and understand how they alter protein function and contribute to Robinow pathogenesis. 2) In our ongoing analysis of the Wnt5a-Ror signaling target Dishevelled (Dvl), it has become essential to separate the different Dvl complexes that mediate different modes of Wnt signaling, including the canonical Wnt/beta-catenin signaling and the noncaonical Wnt5a-Ror signaling. A combination of high-resolution ion exchange and size exclusion chromatography is need to perform these experiments. 3) Lastly, through affinity purification/mass spectrometry (AP/MS), we recently identified several novel binding partners of the Wnt5a-Ror signaling target Kif26b. To investigate how these binding partners interact with Kif26b and participate in Wnt5a-Ror signaling, we need to isolate and analyze the native Kif26b complex from cell and tissue extracts. These experiments also critically depend on the use of the ÄKTA FPLC system. My lab previously inherited an old FPLC system from a senior colleague. However, the old system, now more than 25 years old, has deteriorated to the point that it is no longer in working conditions. It is also no longer serviceable by the manufacturer. Therefore, the purchase of a new FPLC is crucial for the continuation and successful completion of our R35-funded projects. It will likely also benefit other NIH investigators in the department. My home department (Cell Biology and Human Anatomy at UC Davis School of Medicine) is committed to provide space to house the new FPLC system, as well as funds for instrument maintenance.