In Vitro Reconstitution Reveals Major Differences Between Human and Bacterial Cytochrome C Synthases

Overview

Journal Elife

Specialty Biology

Date 2021 May 11

PMID 33973521

Citations 7

Authors

Molly C Sutherland

Deanna L Mendez

Shalon E Babbitt

Dustin E Tillman

Olga Melnikov

Nathan L Tran

Noah T Prizant

Andrea L Collier

Robert G Kranz

Affiliations

Soon will be listed here.

Abstract

Cytochromes c are ubiquitous heme proteins in mitochondria and bacteria, all possessing a CXXCH (CysXxxXxxCysHis) motif with covalently attached heme. We describe the first in vitro reconstitution of cytochrome c biogenesis using purified mitochondrial (HCCS) and bacterial (CcsBA) cytochrome c synthases. We employ apocytochrome c and peptide analogs containing CXXCH as substrates, examining recognition determinants, thioether attachment, and subsequent release and folding of cytochrome c. Peptide analogs reveal very different recognition requirements between HCCS and CcsBA. For HCCS, a minimal 16-mer peptide is required, comprised of CXXCH and adjacent alpha helix 1, yet neither thiol is critical for recognition. For bacterial CcsBA, both thiols and histidine are required, but not alpha helix 1. Heme attached peptide analogs are not released from the HCCS active site; thus, folding is important in the release mechanism. Peptide analogs behave as inhibitors of cytochrome c biogenesis, paving the way for targeted control.

Citing Articles

Structural Insights into Mechanisms Underlying Mitochondrial and Bacterial Cytochrome c Synthases.

Childs P, Lowder E, Mendez D, Babbitt S, Martinie A, Huynh J Biomolecules. 2025; 14(12.

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Helicobacter pylori and Campylobacter jejuni bacterial holocytochrome c synthase structure-function analysis reveals conservation of heme binding.

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Biogenesis of cytochromes and in the electron transport chain of malaria parasites.

Garcia-Guerrero A, Marvin R, Blackwell A, Sigala P bioRxiv. 2024; .

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