# Function and regulatory mechanisms of the Wnt5a-Ror morphogenetic pathway

> **NIH NIH R35** · UNIVERSITY OF CALIFORNIA AT DAVIS · 2024 · $83,438

## Abstract

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.

## Key facts

- **NIH application ID:** 11098144
- **Project number:** 3R35GM144341-03S1
- **Recipient organization:** UNIVERSITY OF CALIFORNIA AT DAVIS
- **Principal Investigator:** Hsin-Yi Henry Ho
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $83,438
- **Award type:** 3
- **Project period:** 2022-02-01 → 2027-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11098144

## Citation

> US National Institutes of Health, RePORTER application 11098144, Function and regulatory mechanisms of the Wnt5a-Ror morphogenetic pathway (3R35GM144341-03S1). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/11098144. Licensed CC0.

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