Dataset Information

Exoproteome and surfaceome of toxigenic Corynebacterium diphtheriae 1737 and their response to iron-restriction and growth on human hemoglobin

ABSTRACT: Diphtheria is a severe upper respiratory disease that can cause fatal infections. It is known to produce diphtheria toxin and a range of other virulence factors, particularly when it encounters low levels of iron at sites of infection. Genomic sequencing of the NCTC13129 reference strain and its relatives responsible for the Eastern European outbreak in the 1990s revealed the presence of pathogenicity islands that contain a number of genes that encode for surface and secreted virulence factors, including pili, heme-acquisition machinery, and antimicrobial-resistance genes. Despite its clinical relevance and use as a model bacterium for research purposes, no proteomic study reported to date has rigorously defined the exoproteome and surfaceome in any tox-producing C. diphtheriae strain or defined how these proteomes change in response to iron limitation. Thus, defining the full complement of secreted (exoproteome) and surface (surfaceome) protein factors produced by C. diphtheriae that enables it to colonize its host promises to provide insight into how it both causes and prevents disease. To gain insight into how it colonizes its host we have identified iron-dependent changes in the exoproteome and surfaceome of C. diphtheriae strain 1737 using a combination of whole-cell fractionation, intact cell surface proteolysis, and quantitative proteomics. We first performed whole-cell fractionation and surface proteomics to quantitatively evaluate the exoproteome and surfaceome under low-iron growth conditions. For each protein we quantified its degree of secretion and surface-exposure by normalizing supernatant and surface digest abundances to fractions derived from the whole-cell lysate. Then, we performed differential abundance on secreted (supernatant) and surface (surface proteolysis) proteins under three different iron conditions that tested both the effect of iron concentration and iron source (e.g. free iron or heme from hemoglobin) on the exoproteome and surfaceome, respectively. All experiments were performed in biological triplicate, for a total of 33 LC-MS/MS runs.

INSTRUMENT(S):

ORGANISM(S): Corynebacterium Diphtheriae Nctc 13129 Bacteria

TISSUE(S): Cell Culture

DISEASE(S): Diphtheria

SUBMITTER: Andrew Goring

LAB HEAD: Joseph Loo

PROVIDER: PXD051674 | Pride | 2025-05-06

REPOSITORIES: Pride

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The Exoproteome and Surfaceome of Toxigenic Corynebacterium diphtheriae 1737 and Its Response to Iron Restriction and Growth on Human Hemoglobin.

Goring Andrew K AK Hale Scott S Dasika Poojita P Chen Yu Y Clubb Robert T RT Loo Joseph A JA

Journal of proteome research 20241218 1

Toxin-producing Corynebacterium diphtheriae strains are the etiological agents of the severe upper respiratory disease, diphtheria. A global phylogenetic analysis revealed that biotype gravis is particularly lethal as it produces diphtheria toxin and a range of other virulence factors, particularly when it encounters low levels of iron at sites of infection. To gain insight into how it colonizes its host, we have identified iron-dependent changes in the exoproteome and surfaceome of C. ...[more]

PMID: 39692319

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Project description:Gene expression along the crypt-villus (C-V) axis was analyzed using cryostat sectioning to isolate fractions representing the crypts (bottom) and villus tops (top). These fractions were used for analyzing gene expression in iron replete Wistar rats (++), iron deficient Wistar rats (low iron), and in iron deficient Wistar rats fed iron for 3 and 6 days (iron-fed). Differences were observed between the crypts and villus tops in the expression of genes associated with Wnt and BNP signaling, cell proliferation and apoptosis, lipid and iron transport and metabolism. Gene expression in villus crypts and tops was also compared between Wistar and Belgrade rats (bb) and Belgrade rats fed iron (iron-fed) particularly as related to iron absorption and metabolism to define the affects of the mutation in DMT1 in the Belgrade rat on the expression of genes related to iron absorption and metabolism and the response to iron feeding. Keywords: iron stress response A colony of Wistar strain rats (++) and Belgrade (bb) rats on a Wistar background maintained in our animal quarters was used for this study. Twelve one-month old ++ rats were separated into 3 groups of four rats each. Group 1 rats (samples 1-8) were fed a normal diet and served as control animals. Group 2 (samples 9-16) and Group 3 rats (samples 17-22) were fed a low iron diet and bled 2-3 ml every three days for 4 weeks and served as iron deficient rats (low-iron). Group 3 rats, of which one rat died during anesthesia while undergoing phlebotomy, were fed iron supplementation in drinking water (50 ÂµM Ferric ammonium citrate) for the three days before sacrifice (iron-fed). Rats were fasted overnight, killed by injection of pentobarbital sodium and a 1-cm duodenal segment 1 cm distal from the pylorus was cryostat sectioned at the right angle to the crypt-villus axis and the sections representing the top one-third of villus (top) and the bottom one-third of the villus were pooled for RNA isolation. For the study of Belgrade rats, eleven bb rats were divided into 3 groups with the 4 rats in Group 1 (samples 23-30) maintained on a normal diet, the 4 rats in Group 2 (samples 31-38) fed iron for 3 days prior to sacrifice, and the 3 rats in Group 3 (samples 39-44) fed iron for 6 days prior to sacrifice. All procedures for the bb rats were the same as described above for the ++ rats. An additional ++ rat (samples 45-46) was similarly analyzed in a separate experiment.

			Action	DRS
	CdLogHigh_Dig1.raw	Raw
	CdLogHigh_Dig2.raw	Raw
	CdLogHigh_Dig3.raw	Raw
	CdLogHigh_Sup1.raw	Raw
	CdLogHigh_Sup2.raw	Raw

Dataset Information

Exoproteome and surfaceome of toxigenic Corynebacterium diphtheriae 1737 and their response to iron-restriction and growth on human hemoglobin

Dataset's files

Publications

The Exoproteome and Surfaceome of Toxigenic <i>Corynebacterium diphtheriae</i> 1737 and Its Response to Iron Restriction and Growth on Human Hemoglobin.

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Dataset Information

Exoproteome and surfaceome of toxigenic Corynebacterium diphtheriae 1737 and their response to iron-restriction and growth on human hemoglobin

Dataset's files

Publications

The Exoproteome and Surfaceome of Toxigenic &lt;i&gt;Corynebacterium diphtheriae&lt;/i&gt; 1737 and Its Response to Iron Restriction and Growth on Human Hemoglobin.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

The Exoproteome and Surfaceome of Toxigenic <i>Corynebacterium diphtheriae</i> 1737 and Its Response to Iron Restriction and Growth on Human Hemoglobin.