# The global regulation of dynamics and structure mediated by single hydride in a family of reductases

> **NIH NIH R01** · UNIVERSITY OF COLORADO DENVER · 2022 · $302,589

## Abstract

PROJECT SUMMARY
 We have discovered that a single hydride induces global changes to both structure and dynamics within
multiple members of an enzyme family, providing a fundamental link between enzyme structure, dynamics, and
allostery that has implications to the entire oxidoreductase superfamily. Specifically, the BLVRB family are
NADPH-dependent reductases present in multiple organisms where they regulate cellular redox through the
reduction of biliverdin-to-bilirubin and a wide array of flavin substrates. While our recent publications have
revealed that coenzyme binding is coupled to global conformational and dynamic changes, we have now
discovered that there are largescale changes coupled to the oxidation state of the coenzyme as far as 23 Å
away. Thus,
structural
catalytic
the central premise of this application is that a coenzyme's hydride is globally coupled to both
and dynamic changes within an enzyme family and that such global coupling is integrally related to
function.
 The novelty here is that we will explicitly determine how a single hydride, i.e., the difference between
NADPH/NADP+, is globally linked (Aim 1) and how this global coupling controls enzyme function (Aim 2).
Further innovation includes the following. First, we have discovered that hydride-coupled networks can be
modulated by mutations directly to the enzyme/coenzyme interface but also to distally coupled sites, which
gives us the unique opportunity to determine the role of these networks in function. Second, we have
discovered that evolutionarily changing residues modulate hydride coupled networks and function, providing
remarkable insight into the evolutionary role of hydride-mediated coupling and function. Evolutionary
differences will therefore be exploited to identify allosteric networks coupled to the oxidative state of the
coenzyme and simultaneously reveal their evolutionary roles in function. Based on our preliminary data that
includes NMR, X-ray crystallographic, and biochemical studies, we hypothesize that the coenzyme oxidation
induces its own conformational change that is further propagated globally through the enzyme in multiple
BLVRB family members (referred to as “insideout” coupling) and that networks coupled to these changes
modulate function (referred to as “outsidein” coupling). We will address this hypothesis through the following:
Aim 1) Determine how a single hydride modulates the global dynamics and structure within the BLVRB
family of enzymes. NMR solution studies using CSPs, relaxation studies, and ensembles methods will be
used to determine how a single hydride imparts its global regulation to dynamics and structure using three
distinct BLVRB family members with both active site and distal differences (human, hyrax, and mosquito).
Aim 2) Determine the functional role of networks coupled to the oxidative state of the coenzyme.
Biochemical and biophysical methods will be used to determine the functional role of hydride-mediated global
regulati...

## Key facts

- **NIH application ID:** 10489331
- **Project number:** 5R01GM139892-02
- **Recipient organization:** UNIVERSITY OF COLORADO DENVER
- **Principal Investigator:** ELAN Z EISENMESSER
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $302,589
- **Award type:** 5
- **Project period:** 2021-09-15 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10489331

## Citation

> US National Institutes of Health, RePORTER application 10489331, The global regulation of dynamics and structure mediated by single hydride in a family of reductases (5R01GM139892-02). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10489331. Licensed CC0.

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