# Evolutionary Origins of Protein Regulation

> **NIH NIH R01** · FLORIDA STATE UNIVERSITY · 2021 · $298,302

## Abstract

Protein regulation is ubiquitous in biology. It plays an essential role in controlling and coordinating a variety
of cellular processes including replication, division and growth. Protein mis-regulation has been implicated
in many human diseases. The emergence of new regulatory strategies often requires protein conformational
heterogeneity. For example, allostery requires the ability to toggle between two (or more) distinct
polypeptide structures. Similarly, a protein's ability to interact with multiple binding partners often requires
sampling a range of conformations. Past experimental studies of individual polypeptides have yielded a
molecular level understanding of many conformation-dependent regulatory processes. Despite this fact, we
know little about how amino acid sequence changes serve to alter a protein's conformational landscape,
such that new regulatory strategies can be realized. The goal of this proposal is to illuminate how
evolutionary trajectories, shaped by natural selection, provide access to new protein conformations, which
facilitate the emergence of novel regulatory mechanisms. As a model system, we will study the evolution of
regulation within the glucokinase (GCK) protein family. GCK is regulated by several mechanisms that
require access to multiple conformations, and GCK is evolutionarily tractable – its orthology is established
and sequences from a range of species are available. We will use ancestral sequence reconstruction to
resurrect ancestral versions of GCK and it's regulatory protein, GKRP. We will perform structural,
biochemical and biophysical analyses of these proteins to illuminate the functional transitions that afford
access to new protein conformations, which facilitate novel modes of regulation. Understanding the
evolutionary origins of GCK's diverse regulatory processes will provide direct insight into the emergence of
physiology and disease, as GCK is the primary glucose sensor in humans and a key determinant of glucose
homeostasis in vertebrates. Fundamental questions that will be addressed in this proposal include: (1) How
does a disordered polypeptide region evolve from an ordered precursor? (2) How do sequence changes
expand the conformational landscape of a protein? (3) What events led to the emergence of allostery? (4)
How does a new regulatory protein-protein interaction arise from two non-interacting ancestral proteins?

## Key facts

- **NIH application ID:** 10200092
- **Project number:** 5R01GM133843-03
- **Recipient organization:** FLORIDA STATE UNIVERSITY
- **Principal Investigator:** Brian Gene Miller
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $298,302
- **Award type:** 5
- **Project period:** 2019-09-20 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10200092

## Citation

> US National Institutes of Health, RePORTER application 10200092, Evolutionary Origins of Protein Regulation (5R01GM133843-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10200092. Licensed CC0.

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