# Chemogenetic control of kinase and phosphatase activity by modulating autoinhibition

> **NIH NIH R21** · STANFORD UNIVERSITY · 2021 · $233,654

## Abstract

ABSTRACT
Signaling enzymes such as kinases and phosphatases control multiple aspects of cellular differentiation and
behavior, and are especially important in transducing signals from cell surface receptors to changes in cell fate
or function. The ability to activate signaling proteins of interest using validated cell-permeable drugs would be
immensely useful for studying the functions of these proteins in cells or animals, and could provide much-needed
control over gene and cell therapies.
Here, we propose a novel method for conferring chemical control over kinases and phosphatases based on
drug-induced displacement of a tethered autoinhibitory domain (AID) from the active site. We will test and
validate this method using the phosphatase calcineurin (CaN) and calcium/calmodulin kinase IV (CaMKIV), two
enzymes that are natively inhibited by an AID and activated by a mechanism involving AID dissociation. In our
method, we will use fused heterodimerizing elements to position the AIDs near the enzyme active site, then use
small-molecule drugs to disrupt this interaction and displace the AIDs from the active site. We will carry out the
following specific aims: (1) Creating drug-activated CaN using a chemically-dissociable autoinhibitory peptide,
(2) Creating drug-activated CaMKIV using a chemically-dissociable autoinhibitory peptide, and (3) Examining
roles of CaN and CaMKIV in IL-2 transcription in T cells using drug-activated proteins.
Our design has several unique and innovative features. The single-chain design should improve reliability and
reduce complexity over multi-component systems. The ability to rationally modulate linker length, heterodimer
affinity, and AID-enzyme affinity provides multiple avenues for construct optimization. Multiple chemically
dissociable interactions are known, allowing for multiplexed drug-controllable proteins. Finally, given that
intramolecular AIDs should be low rather than high affinity, peptide inhibitors can be selected or designed for
signaling enzymes that lack native AIDs. Our method of protein control by drug-induced displacement of an
autoinhibitory domain should thus be uniquely useful, robust, and generalizable.

## Key facts

- **NIH application ID:** 10195182
- **Project number:** 1R21GM141677-01
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Michael Z. Lin
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $233,654
- **Award type:** 1
- **Project period:** 2021-04-01 → 2023-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10195182

## Citation

> US National Institutes of Health, RePORTER application 10195182, Chemogenetic control of kinase and phosphatase activity by modulating autoinhibition (1R21GM141677-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10195182. Licensed CC0.

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