# Functional phosphosignaling in Mtb infection

> **NIH NIH R21** · SEATTLE CHILDREN'S HOSPITAL · 2020 · $279,531

## Abstract

PROJECT SUMMARY
Phosphosignaling coordinates much of the host macrophage’s (M’s) response to infection with
Mycobacterium tuberculosis (Mtb). Surprisingly, only few studies to date have measured global changes in the
phosphoproteome upon infection with Mtb by mass spectrometry, and none in primary human Ms. In
addition, a major challenge in the cell signaling field today is to separate phosphorylation events that are
functional from those that are not. The lack of tools, experimental or computational, to d istinguish functional
from non-functional phosphorylation events is the reason why many current studies of phosphosignaling
largely remain inventories of phosphosites that provide limited biological insight. Combining phospho- and
structural proteomics, we will test the hypothesis that the occurrence of a structural change of a protein upon
phosphorylation is a reliable predictor of functional phosphorylation events. We will test this idea by combining
detailed phosphosite mapping and global protein structure measurements to compare the proteomes of
uninfected to Mtb-infected primary human Ms. We will correlate changes in individual phosphorylation sites
with changes in the proteolytic cleavage pattern as a readout of structural change. Phosphorylation events that
change a protein’s structure will be tested for their function in Mtb infection. We will knock out (or down) the
phosphoprotein and test the effect on bacterial loads and M function. To link effects of knockout to specific
phosphorylation sites, we will complement with the protein bearing phosphoablative and -mimetic mutations
and test for effects on M signaling and Mtb infection. This project will provide a proof-of-principle for an
experimental method to identify functional phosphorylation sites in general, and in particular in the context of
the Mtb-infected M. These data will allow for more meaningful curation of M phosphorylation datasets,
provide a global, functional view of the cell’s remodeling upon infection, highlight the specific immune pathways
triggered by Mtb infection, identify potential new host-directed therapy targets, and identify downstream
effectors that control or exacerbate Mtb infection.

## Key facts

- **NIH application ID:** 10040388
- **Project number:** 1R21AI153766-01
- **Recipient organization:** SEATTLE CHILDREN'S HOSPITAL
- **Principal Investigator:** Christoph Grundner
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $279,531
- **Award type:** 1
- **Project period:** 2020-06-02 → 2022-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10040388

## Citation

> US National Institutes of Health, RePORTER application 10040388, Functional phosphosignaling in Mtb infection (1R21AI153766-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10040388. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*