# Pyruvate Dehydrogenase Assembly, Structure & Function

> **NIH NIH R01** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2020 · $301,263

## Abstract

The overall goal is to fill substantial gaps in our existing knowledge about the medically important Pyruvate
Dehydrogenase Multi-Enzyme Complex (PDHc), and provide the most detailed information possible
regarding its' structure-function relationships. PDHc plays a critical role in cellular energy production and
carbohydrate metabolism, with disruptions or malfunctions in it associated with numerous pathologies.
Bacterial PDHc was also recently recognized as the target for an antibiotic, TPBC, currently in hospital use that
is highly effective against emerging health threats including MRSA (methicillin resistant staphylococcus aureus)
and VRE (vancomycin resistant enterococci), but its inhibition mechanism is unknown. We will focus on
obtaining details for the overall assembly and unusual catalytic process employed by PDHc's, and in particular,
for the bacterial PDHc from E. coli. We will also examine differences between the E. coli and human versions.
The overall reaction, converting pyruvate to acetyl-CoA while releasing NADH, is carried out by at least three
different enzymes with multiple copies of each in large complexes having different architectures. The long-term
goal is to understand structure-function relationships in the intact, octahedral, PDHc complex from E. coli and
compare them with the icosahedral PDHc complex from humans. We will determine and analyze specific
interactions required for governing the assembly of, and catalysis within, the multi-enzyme complex. Unlike
most biochemical pathways operating by simple diffusion of substrates between isolated enzymes, PDHc
employs complex architectures utilizing substrate channeling involving long and flexible swinging-arms to
“hand-deliver” intermediates between the multiple (at least 60) E1, E2, and E3 enzymatic component active
sites. We successfully completed structural analyses of the E1 and E3 components, the E2 core, and some
key sub-complexes. This work forms the basis of the current proposal to obtain details for some key missing
interactions, and combine them with our earlier data to assess the entire assembly. We will determine and
analyze crystal structures for the few missing binary complexes, and correlate the results with overall biological
function. We will also provide structure/function information on protein-protein interactions governing
assembly/catalysis in the human, icosahedral, PDHc complex, which differs significantly from the octahedral E.
coli version. In addition, we will structurally determine/analyze human PDHc components and their binary
complexes to assess if differences relative to E. coli PDHc may be exploitable for anti-bacterial drug
development, and to determine the structural basis for diseases associated with single residue mutations in
human PDHc proteins. We will then put everything together, by analyzing the entire E. coli PDHc structure
either by cryo-EM or x-ray methods. Structural studies on isolated proteins or protein-protein complexes wil...

## Key facts

- **NIH application ID:** 9942418
- **Project number:** 5R01GM121469-04
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** WILLIAM F FUREY
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $301,263
- **Award type:** 5
- **Project period:** 2017-07-01 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9942418

## Citation

> US National Institutes of Health, RePORTER application 9942418, Pyruvate Dehydrogenase Assembly, Structure & Function (5R01GM121469-04). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/9942418. Licensed CC0.

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