Membrane proteome of the green sulfur bacterium Chlorobium tepidum (syn. Chlorobaculum tepidum) analyzed by gel-based and gel-free methods

Kouyianou, Kalliopi; Aivaliotis, Michalis; Gevaert, Kris; Karas, Michael; Tsiotis, Georgios

doi:10.1007/s11120-010-9544-x

Cited by 4 publications

(4 citation statements)

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“…We have also demonstrated higher levels of protein overlap for gel-free analysis of biological replicates of other bacterial species ( M. haemolytica and P. multocida ) and believe that the lower overlap observed for Y. ruckeri is a species-specific phenomenon. Clearly, the use of these complementary approaches resulted in improved overall coverage of the outer membrane proteome and was consistent with previous work in other bacterial species [44,[61], [62], [63],83,84].…”

Section: Discussionsupporting

confidence: 87%

Comparative bioinformatic and proteomic approaches to evaluate the outer membrane proteome of the fish pathogen Yersinia ruckeri

Ormsby

Grahame

Burchmore

et al. 2019

Journal of Proteomics

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Yersinia ruckeri is the aetiological agent of enteric redmouth (ERM) disease and is responsible for significant economic losses in farmed salmonids. Enteric redmouth disease is associated primarily with rainbow trout ( Oncorhynchus mykiss, Walbaum) but its incidence in Atlantic salmon ( Salmo salar ) is increasing. Outer membrane proteins (OMPs) of Gram-negative bacteria are located at the host-pathogen interface and play important roles in virulence. The outer membrane of Y. ruckeri is poorly characterised and little is known about its composition and the roles of individual OMPs in virulence. Here, we employed a bioinformatic pipeline to first predict the OMP composition of Y. ruckeri . Comparative proteomic approaches were subsequently used to identify those proteins expressed in vitro in eight representative isolates recovered from Atlantic salmon and rainbow trout. One hundred and forty-one OMPs were predicted from four Y. ruckeri genomes and 77 of these were identified in three or more genomes and were considered as “core” proteins. Gel-free and gel-based proteomic approaches together identified 65 OMPs in a single reference isolate and subsequent gel-free analysis identified 64 OMPs in the eight Atlantic salmon and rainbow trout isolates. Together, our gel-free and gel-based proteomic analyses identified 84 unique OMPs in Y. ruckeri . Significance Yersinia ruckeri is an important pathogen of Atlantic salmon and rainbow trout and is of major economic significance to the aquaculture industry worldwide. Disease outbreaks are becoming more problematic in Atlantic salmon and there is an urgent need to investigate in further detail the cell-surface (outer membrane) composition of strains infecting each of these host species. Currently, the outer membrane of Y. ruckeri is poorly characterised and very little is known about the OMP composition of strains infecting each of these salmonid species. This study represents the most comprehensive comparative outer membrane proteomic analysis of Y. ruckeri to date, encompassing isolates of different biotypes, serotypes, OMP-types and hosts of origin and provides insights into the potential roles of these diverse proteins in host-pathogen interactions. The study has identified key OMPs likely to be involved in disease pathogenesis and makes a significant contribution to furthering our understanding of the cell-surface composition of this important fish pathogen that will be relevant to the development of improved vaccines and therapeutics.

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Section: Discussionsupporting

confidence: 87%

Comparative bioinformatic and proteomic approaches to evaluate the outer membrane proteome of the fish pathogen Yersinia ruckeri

Ormsby

Grahame

Burchmore

et al. 2019

Journal of Proteomics

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“…Similar results were also obtained from mixotrophic growth with pyruvate or acetate (data not shown). Moreover, SDH/fumarate reductase-like proteins encoded by CT2040 -2042, not CT2266 -2268, were also reported by recent proteomic studies (27).…”

Section: Labeled Bchls In the Mixotrophic Cultures Withmentioning

confidence: 63%

“…3A also indicates that the transcript level of aclAB is ϳ6 -10-fold higher than gltA. The CS protein has been previously reported by the proteomic analysis of C. tepidum (27,28), and the studies by Hosoya-Matsuda et al (28) suggested that CS is inactivated under conditions when ACL is activated. Consistent with their working hypothesis, the enzymatic activity of ACL, not that of CS, can be detected in cell extracts of autotrophic and mixotrophic growth cultures (Fig.…”

Section: Labeled Bchls In the Mixotrophic Cultures Withmentioning

confidence: 87%

Both Forward and Reverse TCA Cycles Operate in Green Sulfur Bacteria

Tang

Blankenship

2010

Journal of Biological Chemistry

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The anoxygenic green sulfur bacteria (GSBs) assimilate CO 2 autotrophically through the reductive (reverse) tricarboxylic acid (RTCA) cycle. Some organic carbon sources, such as acetate and pyruvate, can be assimilated during the phototrophic growth of the GSBs, in the presence of CO 2 or HCO 3 ؊ . It has not been established why the inorganic carbonis required for incorporating organic carbon for growth and how the organic carbons are assimilated. In this report, we probed carbon flux during autotrophic and mixotrophic growth of the GSB Chlorobaculum tepidum. Our data indicate the following: (a) the RTCA cycle is active during autotrophic and mixotrophic growth; (b) the flux from pyruvate to acetyl-CoA is very low and acetyl-CoA is synthesized through the RTCA cycle and acetate assimilation; (c) pyruvate is largely assimilated through the RTCA cycle; and (d) acetate can be assimilated via both of the RTCA as well as the oxidative (forward) TCA (OTCA) cycle. The OTCA cycle revealed herein may explain better cell growth during mixotrophic growth with acetate, as energy is generated through the OTCA cycle. Furthermore, the genes specific for the OTCA cycle are either absent or down-regulated during phototrophic growth, implying that the OTCA cycle is not complete, and CO 2 is required for the RTCA cycle to produce metabolites in the TCA cycle. Moreover, CO 2 is essential for assimilating acetate and pyruvate through the CO 2 -anaplerotic pathway and pyruvate synthesis from acetyl-CoA.Chlorobaculum tepidum (formerly Chlorobium tepidum) (1) is a phototrophic green sulfur bacterium (GSB) 2 that fixes carbon photoautotrophically through the reductive (reverse) tricarboxylic acid (RTCA) cycle (see Fig. 1A) (2). The RTCA cycle, first reported in a green sulfur bacterium Chlorobium thiosulfatophilum (3), is essentially the reversal of the oxidative (forward) tricarboxylic acid (OTCA) cycle. Four enzymes have been recruited for the RTCA cycle to catalyze the reverse reaction of four steps in the OTCA cycle (3); pyruvate:ferredoxin (Fd) oxidoreductase (acetyl-CoA ϩ CO 2 ϩ 2Fd red ϩ 2H ϩ º pyruvate ϩ CoA ϩ 2Fd ox ), ATP citrate lyase (ACL, acetyl-CoA ϩ oxaloacetate ϩ ADP ϩ P i º citrate ϩ CoA ϩ ATP), ␣-ketoglutarate:ferredoxin oxidoreductase (succinyl-CoA ϩ CO 2 ϩ 2Fd red ϩ 2H ϩ º ␣-ketoglutartate ϩ CoA ϩ 2Fd ox ) and fumarare reductase (succinate ϩ acceptor º fumarate ϩ reduced acceptor). Moreover, several GSBs, C. tepidum included, are potential mixotrophs that use organic carbon sources for producing biomass in the presence of CO 2 or HCO 3 Ϫ (2, 4, 5). Fluoroacetate (FAc) is known to be a metabolic poison that is highly toxic to mammals. As a structural analog of acetate, the metabolic pathway of FAc has been suggested to be similar to that of acetate, and the toxicity of FAc can be reduced by acetate (6). FAc is known to be an inhibitor for the OTCA cycle (Fig. 1B). Like acetate, FAc is converted into fluoroacetyl-CoA (7), which condenses with oxaloacetate (OAA) to produce (Ϫ)- erythro-(2R,3R)-2-fluorocitrate (2-FC)...

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“…Qualitative and quantitative mass spectrometry-based proteomics have become a routine tool to examine proteome remodelling in different cellular states [15]. The whole proteome of C. tepidum is extensively studied by gel-based and gel-free methods [16][17][18][19][20][21][22]. Some reports have employed sample separation by 2D-IEF/SDS-PAGE and 2D-BN/SDS-PAGE allowing the identification of about 625 different [16,17,20].…”

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confidence: 99%

Insights into the sulfur metabolism of Chlorobaculum tepidum by label‐free quantitative proteomics

et al. 2023

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Chlorobaculum tepidum is an anaerobic green sulfur bacterium which oxidizes sulfide, elemental sulfur, and thiosulfate for photosynthetic growth. It can also oxidize sulfide to produce extracellular S 0 globules, which can be further oxidized to sulfate and used as an electron donor. Here, we performed label-free quantitative proteomics on total cell lysates prepared from different metabolic states, including a sulfur production state (10 h post-incubation [PI]), the beginning of sulfur consumption (20 h PI), and the end of sulfur consumption (40 h PI), respectively. We observed an increased abundance of the sulfide:quinone oxidoreductase (Sqr) proteins in 10 h PI indicating a sulfur production state. The periplasmic thiosulfate-oxidizing Sox enzymes and the dissimilatory sulfite reductase (Dsr) subunits showed an increased abundance in 20 h PI, corresponding to the sulfur-consuming state. In addition, we found that the abundance of the heterodisulfide-reductase and the sulfhydrogenase operons was influenced by electron donor availability and may be associated with sulfur metabolism. Further, we isolated and analyzed the extracellular sulfur globules in the different metabolic states to study their morphology and the sulfur cluster composition, yielding 58 previously uncharacterized proteins in purified globules. Our results show that C. tepidum regulates the cellular levels of enzymes involved in sulfur metabolism in response to the availability of reduced sulfur compounds. K E Y W O R D Selemental sulfur, green sulfur bacteria, label-free quantitative, proteomics, sulfur metabolism INTRODUCTIONChlrobaculum (Cba) tepidum is a strictly anaerobic, photosynthetic bacterium that predominantly oxidizes reduced sulfur compounds for photoautotrophic growth [1]. C. tepidum was isolated from a hot spring in New Zealand and preferentially oxidizes sulfide to elemental sulfur,

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Membrane proteome of the green sulfur bacterium Chlorobium tepidum (syn. Chlorobaculum tepidum) analyzed by gel-based and gel-free methods

Cited by 4 publications

References 44 publications

Comparative bioinformatic and proteomic approaches to evaluate the outer membrane proteome of the fish pathogen Yersinia ruckeri

Comparative bioinformatic and proteomic approaches to evaluate the outer membrane proteome of the fish pathogen Yersinia ruckeri

Both Forward and Reverse TCA Cycles Operate in Green Sulfur Bacteria

Insights into the sulfur metabolism of Chlorobaculum tepidum by label‐free quantitative proteomics

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