# Chemical Approaches to Cell Signaling Enzymes

> **NIH NIH R01** · BRIGHAM AND WOMEN'S HOSPITAL · 2022 · $345,002

## Abstract

Abstract
This is a competing renewal application of our longstanding R01 Grant CA74305 that
concerns the development and application of chemical approaches to enhance the
understanding of cell signaling pathways and the attachment, removal, and function of
protein post-translational modifications (PTMs). In the next cycle, we plan to investigate
how ubiquitination regulates two key cancer-related signaling enzymes, PTEN and Akt,
how several cancer-connected E3 ubiquitin (Ub) ligases target and catalyze Ub transfer
to their protein substrates, and how mTORC2 phosphorylates its major substrate Akt.
PTEN is one of the most commonly mutated tumor suppressor genes and catalyzes the
conversion of phospholipid phosphatidylinositol-3,4,5-triphosphate (PIP3) to PIP2 and in
this way antagonizes the action of the PI3-kinase/Akt oncogenic signaling pathway. Akt
is a protein Ser/Thr kinase that is activated by PIP3 and drives tumor formation. Akt is an
intensively studied cancer therapeutic target. E3 Ub ligases are a large family (>600
members) of enzymes that target Akt, PTEN, and thousands of cellular substrates for
Lys ubiquitination. Our research program will continue to develop and employ protein
semisynthetic methods including ubiquitin hydrazide installation to generate mimics of
mono-ubiquitinated proteins and covalent E3-Ub-substrate ternary complexes. Our three
specific aims are: 1. Elucidate the basis of PTEN and Akt regulation by Lys
ubiquitination; 2. Delineate the catalytic mechanisms and protein substrate recognition of
key E3 ubiquitin ligases; 3. Determine the molecular basis of mTORC2 mediated-
phosphorylation of Akt. With site-specifically modified signaling proteins in hand, we will
integrate kinetic assays, structural analysis, binding measurements, and cell-based
studies to clarify key regulatory and signaling features. Upon completion of this research
effort, we will broaden the knowledge of how Lys ubiquitination targets critical cancer-
related proteins and influences their functions. We will also deepen our understanding
of how the multisubunit mTORC2 complex is able to activate Akt. Moreover, these
studies should pave the way for new therapeutic strategies to combat pathway
dysregulation in cancer. This research program will also enable the training of the next
generation of biochemical investigators with an interest in cancer.

## Key facts

- **NIH application ID:** 10444002
- **Project number:** 2R01CA074305-28
- **Recipient organization:** BRIGHAM AND WOMEN'S HOSPITAL
- **Principal Investigator:** PHILIP A COLE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $345,002
- **Award type:** 2
- **Project period:** 1997-04-01 → 2027-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10444002

## Citation

> US National Institutes of Health, RePORTER application 10444002, Chemical Approaches to Cell Signaling Enzymes (2R01CA074305-28). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10444002. Licensed CC0.

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