# Regulation of beta2-adrenergic receptor signaling by post-translational modifications

> **NIH NIH R01** · MEDICAL COLLEGE OF WISCONSIN · 2020 · $338,800

## Abstract

PROJECT SUMMARY/ABSTRACT
G-protein-coupled receptors (GPCRs) are a large and diverse class of cell surface receptors responsible for
regulating nearly every physiological process in the human body. Upon binding to the extracellular surface of
GPCRs, ligands trigger conformational changes that lead to binding and activating signaling proteins at the
cytoplasmic surface of the receptor. Many GPCRs signal through several pathways that include different
heterotrimeric G-protein subtypes as well as arrestins, and “biased” ligands differentially modulate receptor
signaling through these various pathways. GPCRs rely on a high degree of conformational flexibility to achieve
this signaling complexity, and one of the major goals in the field of structure-based drug design is to identify the
conformations that generate a particular signaling profile. Significant progress toward this goal has been made,
yet relatively little is known about the effects of endogenous modulators, including protein-lipid interactions, post-
translational modifications (PTMs), and accessory proteins, on GPCR structure and dynamics. The effect of
endogenous modulators on the conformational landscape and the interplay of endogenous modulators with
potential drugs need to be characterized to achieve the objectives of rational drug design, and this is a major
long-term goal of my laboratory. In this project, we will focus on two PTMs of the β2-adrenergic receptor (β2AR)—
palmitoylation and glycosylation—with the goal of identifying the structural and dynamical basis for their
regulation of β2AR signaling. A complementary combination of continuous-wave and pulsed EPR techniques,
mass spectrometry, and functional assays will be used in these studies. Importantly, cutting-edge variable-
pressure EPR technology will provide unique mechanistic insights by mapping sparsely populated regions of the
conformational landscape. First, we will map the conformational landscape at the cytoplasmic surface in the
reference state of the receptor, defined herein as the natively glycosylated and palmitoylated receptor embedded
in a lipid bilayer. Then, we will perform experiments with the PTMs removed, one at a time, to identify the
allosteric effects of palmitoylation and glycosylation on the cytoplasmic surface. Finally, we will determine the
local effects of glycosylation on the ligand-binding pocket at the extracellular surface and of palmitoylation on
helix 8 and the C-terminal tail. The results will provide insight into the understudied yet critical role of these PTMs
as regulators of β2AR signaling, and because the β2AR is a prototypical member of the GPCR superfamily, we
anticipate the results to be applicable to other GPCRs with similar PTMs. In detailing the effects of endogenous
modulators on the conformational landscape, our results will increase researchers' ability to rationally design
drugs to achieve the desired therapeutic effect by allowing the in vivo interplay of endogenous modulato...

## Key facts

- **NIH application ID:** 10019579
- **Project number:** 5R01GM135581-02
- **Recipient organization:** MEDICAL COLLEGE OF WISCONSIN
- **Principal Investigator:** MICHAEL TAYLOR LERCH
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $338,800
- **Award type:** 5
- **Project period:** 2019-09-20 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10019579

## Citation

> US National Institutes of Health, RePORTER application 10019579, Regulation of beta2-adrenergic receptor signaling by post-translational modifications (5R01GM135581-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10019579. Licensed CC0.

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