Global role of the membrane protease LonB in Archaea: Potential protease targets revealed by quantitative proteome analysis of a lonB mutant in Haloferax volcanii

Cerletti, Micaela; Paggi, Roberto A.; Guevara, Carina Ramallo; Poetsch, Ansgar; Castro, Rosana Esther de

doi:10.1016/j.jprot.2015.03.016

Cited by 26 publications

(41 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, proteomics is naturally the method of choice when it comes to the discovery of protease targets and proteolytic processing events. In our previous works with microorganisms, Lon target candidates could be identified by common quantitative shotgun proteomics [102] and more sophisticated proteome turnover analysis [103]. There is no reason to believe why similar studies could not be executed in the future for Brucella to illuminate such aspects of protease function.…”

Section: Proteases-a Yet Untouched Topic In Proteomics For Brucellamentioning

confidence: 99%

Proteomics of Brucella

Poetsch

Marchesini

2020

Proteomes

Self Cite

View full text Add to dashboard Cite

Brucella spp. are Gram negative intracellular bacteria responsible for brucellosis, a worldwide distributed zoonosis. A prominent aspect of the Brucella life cycle is its ability to invade, survive and multiply within host cells. Comprehensive approaches, such as proteomics, have aided in unravelling the molecular mechanisms underlying Brucella pathogenesis. Technological and methodological advancements such as increased instrument performance and multiplexed quantification have broadened the range of proteome studies, enabling new and improved analyses, providing deeper and more accurate proteome coverage. Indeed, proteomics has demonstrated its contribution to key research questions in Brucella biology, i.e., immunodominant proteins, host-cell interaction, stress response, antibiotic targets and resistance, protein secretion. Here, we review the proteomics of Brucella with a focus on more recent works and novel findings, ranging from reconfiguration of the intracellular bacterial proteome and studies on proteomic profiles of Brucella infected tissues, to the identification of Brucella extracellular proteins with putative roles in cell signaling and pathogenesis. In conclusion, proteomics has yielded copious new candidates and hypotheses that require future verification. It is expected that proteomics will continue to be an invaluable tool for Brucella and applications will further extend to the currently ill-explored aspects including, among others, protein processing and post-translational modification.

show abstract

Section: Proteases-a Yet Untouched Topic In Proteomics For Brucellamentioning

confidence: 99%

Proteomics of Brucella

Poetsch

Marchesini

2020

Proteomes

Self Cite

View full text Add to dashboard Cite

show abstract

“…The whole proteome composition of several neutrophilic haloarchaea has been reported. This includes the high‐throughput proteome characterization of H. volcanii parental strain and protease mutants like the proteasome, the LonB, and the RhoII proteases; Haloarcula marismortui during mid‐log and late‐log phases; and Halobacterium salinarum under different salt concentrations . Meanwhile, in the haloalkaliphilic group only the cytoplasmic proteome of one member ( Natronomonas pharaonis ) has been examined .…”

mentioning

confidence: 99%

“…The clarified cell extract supernatants were precipitated with 1 volume of acetone (O/N, 4 °C) and washed with acetone three times. Protein samples were processed, digested with trypsin, and subjected to LC‐ESI‐MS/MS using a nanoACQUITY gradient UPLC pump system (Waters) and an LTQ Orbitrap Elite mass spectrometer as described in Cerletti et al …”

mentioning

confidence: 99%

Proteomic Study of the Exponential–Stationary Growth Phase Transition in the Haloarchaea Natrialba magadii and Haloferax volcanii

et al. 2018

Self Cite

View full text Add to dashboard Cite

The dynamic changes that take place along the phases of microbial growth (lag, exponential, stationary, and death) have been widely studied in bacteria at the molecular and cellular levels, but little is known for archaea. In this study, a high-throughput approach was used to analyze and compare the proteomes of two haloarchaea during exponential and stationary growth: the neutrophilic Haloferax volcanii and the alkaliphilic Natrialba magadii. Almost 2000 proteins were identified in each species (≈50% of the predicted proteome). Among them, 532 and 432 were found to be differential between growth phases in H. volcanii and N. magadii, respectively. Changes upon entrance into stationary phase included an overall increase in proteins involved in the transport of small molecules and ions, stress response, and fatty acid catabolism. Proteins related to genetic processes and cell division showed a notorious decrease in amount. The data reported in this study not only contributes to our understanding of the exponential-stationary growth phase transition in extremophilic archaea but also provides the first comprehensive analysis of the proteome composition of N. magadii. The MS proteomics data have been deposited in the ProteomeXchange Consortium with the dataset identifier JPST000395.

show abstract

“…After washing (with destaining solution) and drying the gel pieces in a SpeedVac, trypsin (porcine, sequencing grade, Promega) solution (12.5 ng ml -1 in 25 mM ammonium bicarbonate, pH 8.6) was added. Protein digestion and peptide elution was performed according to Cerletti et al 18 . The extracted peptides were dried using a SpeedVac and stored at -20 °C.…”

Section: In-gel Tryptic Digestionmentioning

confidence: 99%

“…In the case of processive proteases, accumulation of a particular protein in a protease defective strain with respect to the control strain would be an evidence of that protein being a specific substrate. This method was applied by our group to identify candidate targets of the LonB protease in H. volcanii 18 . On the other hand, the situation for regulatory enzymes, such as Rho, is different.…”

Section: Introductionmentioning

confidence: 99%

Haloferax volcanii Proteome Response to Deletion of a Rhomboid Protease Gene

et al. 2018

Self Cite

View full text Add to dashboard Cite

Rhomboids are conserved intramembrane serine proteases involved in cell signaling processes. Their role in prokaryotes is scarcely known and remains to be investigated in Archaea. We previously constructed a rhomboid homologue deletion mutant (ΔrhoII) in Haloferax volcanii, which showed reduced motility, increased novobiocin sensitivity, and an N- glycosylation defect. To address the impact of rhoII deletion on H. volcanii physiology, the proteomes of mutant and parental strains were compared by shotgun proteomics. A total of 1847 proteins were identified (45.8% of H. volcanii predicted proteome), from which 103 differed in amount. Additionally, the mutant strain evidenced 99 proteins with altered electrophoretic migration, which suggested differential post-translational processing/modification. Integral membrane proteins that evidenced variations in concentration, electrophoretic migration, or semitryptic cleavage in the mutant were considered as potential RhoII targets. These included a PrsW protease homologue (which was less stable in the mutant strain), a predicted halocyanin, and six integral membrane proteins potentially related to the mutant glycosylation (S-layer glycoprotein, Agl15) and cell adhesion/motility (flagellin1, HVO_1153, PilA1, and PibD) defects. This study investigated for the first time the impact of a rhomboid protease on the whole proteome of an organism.

show abstract

Global role of the membrane protease LonB in Archaea: Potential protease targets revealed by quantitative proteome analysis of a lonB mutant in Haloferax volcanii

Cited by 26 publications

References 68 publications

Proteomics of Brucella

Proteomics of Brucella

Proteomic Study of the Exponential–Stationary Growth Phase Transition in the Haloarchaea Natrialba magadii and Haloferax volcanii

Haloferax volcanii Proteome Response to Deletion of a Rhomboid Protease Gene

Contact Info

Product

Resources

About