Reference map and comparative proteomic analysis of Neisseria gonorrhoeae displaying high resistance against spectinomycin

2014 ◽  
Vol 63 (3) ◽  
pp. 371-385 ◽  
Author(s):  
Sunanta Nabu ◽  
Ratana Lawung ◽  
Patcharee Isarankura-Na-Ayudhya ◽  
Chartchalerm Isarankura-Na-Ayudhya ◽  
Sittiruk Roytrakul ◽  
...  
Keyword(s):  
Energy Metabolism ◽  
Neisseria Gonorrhoeae ◽  
Protein Expression ◽  
Proteomic Analysis ◽  
Cell Envelope ◽  
Expression Profiles ◽  
Compensatory Mechanisms ◽  
Two Dimensional Gel Electrophoresis ◽  
Comparative Proteomic Analysis ◽  
Reference Map

A proteome reference map of Neisseria gonorrhoeae was successfully established using two-dimensional gel electrophoresis in conjunction with matrix-assisted laser desorption ionization–time of flight mass spectrometry. This map was further applied to compare protein expression profiles of high-level spectinomycin-resistant (clinical isolate) and -susceptible (reference strain) N. gonorrhoeae following treatment with subminimal inhibitory concentrations (subMICs) of spectinomycin. Approximately 200 protein spots were visualized by Coomassie brilliant blue G-250 staining and 66 spots representing 58 unique proteins were subsequently identified. Most of the identified proteins were analysed as cytoplasmic proteins and belonged to the class of energy metabolism. Comparative proteomic analysis of whole protein expression of susceptible and resistant gonococci showed up to 96 % similarity while eight proteins were found to be differentially expressed in the resistant strain. In the presence of subMICs of spectinomycin, it was found that 50S ribosomal protein L7/L12, an essential component for ribosomal translocation, was upregulated in both strains, ranging from 1.5- to 3.5-fold, suggesting compensatory mechanisms of N. gonorrhoeae in response to antibiotic that inhibits protein synthesis. Moreover, the differential expression of proteins involved in energy metabolism, amino acid biosynthesis, and the cell envelope was noticeably detected, indicating significant cellular responses and adaptation against antibiotic stress. Such knowledge provides valuable data, not only fundamental proteomic data, but also knowledge of the mode of action of antibiotic and secondary target proteins implicated in adaptation and compensatory mechanisms.

scholarly journals Impact of Spaceflight and Artificial Gravity on the Mouse Retina: Biochemical and Proteomic Analysis

2018 ◽  
Vol 19 (9) ◽  
pp. 2546 ◽  
Author(s):  
Xiao Mao ◽  
Stephanie Byrum ◽  
Nina Nishiyama ◽  
Michael Pecaut ◽  
Vijayalakshmi Sridharan ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Visual Acuity ◽  
Protein Expression ◽  
Proteomic Analysis ◽  
Vascular Endothelial Cells ◽  
Expression Profiles ◽  
Artificial Gravity ◽  
Ocular Tissue ◽  
Mouse Retina ◽  
Vascular Endothelial

Astronauts are reported to have experienced some impairment in visual acuity during their mission on the International Space Station (ISS) and after they returned to Earth. There is emerging evidence that changes in vision may involve alterations in ocular structure and function. To investigate possible mechanisms, changes in protein expression profiles and oxidative stress-associated apoptosis were examined in mouse ocular tissue after spaceflight. Nine-week-old male C57BL/6 mice (n = 12) were launched from the Kennedy Space Center on a SpaceX rocket to the ISS for a 35-day mission. The animals were housed in the mouse Habitat Cage Unit (HCU) in the Japan Aerospace Exploration Agency (JAXA) “Kibo” facility on the ISS. The flight mice lived either under an ambient microgravity condition (µg) or in a centrifugal habitat unit that produced 1 g artificial gravity (µg + 1 g). Habitat control (HC) and vivarium control mice lived on Earth in HCUs or normal vivarium cages, respectively. Quantitative assessment of ocular tissue demonstrated that the µg group induced significant apoptosis in the retina vascular endothelial cells compared to all other groups (p < 0.05) that was 64% greater than that in the HC group. Proteomic analysis showed that many key pathways responsible for cell death, cell repair, inflammation, and metabolic stress were significantly altered in µg mice compared to HC animals. Additionally, there were more significant changes in regulated protein expression in the µg group relative to that in the µg + 1 g group. These data provide evidence that spaceflight induces retinal apoptosis of vascular endothelial cells and changes in retinal protein expression related to cellular structure, immune response and metabolic function, and that artificial gravity (AG) provides some protection against these changes. These retinal cellular responses may affect blood–retinal barrier (BRB) integrity, visual acuity, and impact the potential risk of developing late retinal degeneration.

Protein expression profiles in pediatric multiple sclerosis: potential biomarkers

2009 ◽  
Vol 15 (4) ◽  
pp. 455-464 ◽  
Author(s):  
KN Rithidech ◽  
L Honikel ◽  
M Milazzo ◽  
D Madigan ◽  
R Troxell ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Mass Spectrometry ◽  
Protein Expression ◽  
Expression Profiles ◽  
Low Frequency ◽  
Complement Factor ◽  
Protein Biomarkers ◽  
Two Dimensional Gel Electrophoresis ◽  
Pediatric Multiple Sclerosis ◽  
Proteomic Approach

The diagnosis of pediatric multiple sclerosis (MS) is challenging due to its low frequency and the overlap with other acquired childhood demyelinating disorders of the central nervous system. To identify potential protein biomarkers which could facilitate the diagnosis, we used two-dimensional gel electrophoresis (2-DE) in combination with mass spectrometry to identify proteins associated with pediatric MS. Plasma samples from nine children with MS and nine healthy subjects, matched in aggregate by age and gender, were analyzed for differences in their patterns of protein expression. We found 12 proteins that were significantly up regulated in the pediatric MS group: alpha-1-acid-glycoprotein 1, alpha-1-B-glycoprotein, transthyretin, apoliprotein-C-III, serum amyloid P component, complement factor-I, clusterin, gelsolin, hemopexin, kininogen-1, hCG1993037-isoform, and vitamin D-binding protein. These results show that 2-DE in combination with mass spectrometry is a highly sensitive technique for the identification of blood-based biomarkers. This proteomic approach could lead to a new panel of diagnostic and prognostic markers in pediatric MS.

Comparative proteomic analysis on human L-02 liver cells treated with varying concentrations of trichloroethylene

2007 ◽  
Vol 23 (2) ◽  
pp. 91-101 ◽  
Author(s):  
Jianjun Liu ◽  
Haiyan Huang ◽  
Xiumei Xing ◽  
Renrong Xi ◽  
Zhixiong Zhuang ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Underlying Mechanism ◽  
Liver Cells ◽  
Control Group ◽  
Two Dimensional Gel Electrophoresis ◽  
Comparative Proteomic Analysis ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Peptide Mass ◽  
Tof Ms

To determine the differential proteomic expressions in human L-02 liver cells induced by varying concentrations of trichloroethylene (TCE), comparative proteomic analysis was performed on human L-02 liver cells which were treated with varying concentrations of TCE. According to the result of MTT test, we designed four different groups, in which the cells were treated with 0 μM (control group), 3, 10 or 40 μM TCE for 24 h, respectively. Comparative analysis of approximately 800 spots resolved by two-dimensional gel electrophoresis (2DE) in the soluble proteomes of L-02 cells from the four different groups resulted in 10 differential proteins. To identify the differential spots, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) was carried out; if the results from the tool were insufficient, tandem MS (MALDI-TOF-TOF-MS) was then performed. The raw data of peptide mass fingerprints (PMFs) and MS/MS spectra were searched against the IPI human data base for exact matches. Then western blot was employed to verify the result of proteomic analysis, the following result confirmed that the results of proteomic analysis were reliable. These results might provide an insight into the underlying mechanism of TCE intoxication and find biological markers for diagnosis and therapy of TCE-induced diseases.

scholarly journals Protein Expression Profiles in an Endosymbiotic Cyanobacterium Revealed by a Proteomic Approach

2006 ◽  
Vol 19 (11) ◽  
pp. 1251-1261 ◽  
Author(s):  
Martin Ekman ◽  
Petter Tollbäck ◽  
Johan Klint ◽  
Birgitta Bergman
Keyword(s):  
Mass Spectrometry ◽  
Protein Expression ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Calvin Cycle ◽  
Pentose Phosphate ◽  
Free Living ◽  
Two Dimensional Gel Electrophoresis ◽  
Proteomic Approach ◽  
Nostoc Strain

Molecular mechanisms behind adaptations in the cyano-bacterium (Nostoc sp.) to a life in endosymbiosis with plants are still not clarified, nor are the interactions between the partners. To get further insights, the proteome of a Nostoc strain, freshly isolated from the symbiotic gland tissue of the angiosperm Gunnera manicata Linden, was analyzed and compared with the proteome of the same strain when free-living. Extracted proteins were separated by two-dimensional gel electrophoresis and were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry combined with tandem mass spectrometry. Even when the higher percentage of differentiated cells (heterocysts) in symbiosis was compensated for, the majority of the proteins detected in the symbiotic cyanobacteria were present in the free-living counterpart, indicating that most cellular processes were common for both stages. However, differential expression profiling revealed a significant number of proteins to be down-regulated or missing in the symbiotic stage, while others were more abundant or only expressed in symbiosis. The differential protein expression was primarily connected to i) cell envelope-associated processes, including proteins involved in exopolysaccharide synthesis and surface and membrane associated proteins, ii) to changes in growth and metabolic activities (C and N), including upregulation of nitrogenase and proteins involved in the oxidative pentose phosphate pathway and downregu-lation of Calvin cycle enzymes, and iii) to the dark, micro-aerobic conditions offered inside the Gunnera gland cells, including changes in relative phycobiliprotein concentrations. This is the first comprehensive analysis of proteins in the symbiotic state.

Comparative Proteomics of Commensal and Pathogenic Strains of Escherichia coli

2020 ◽  
Vol 27 (11) ◽  
pp. 1171-1177
Author(s):  
Neelja Singhal ◽  
Divakar Sharma ◽  
Manish Kumar ◽  
Deepa Bisht ◽  
Jugsharan Singh Virdi
Keyword(s):  
Escherichia Coli ◽  
Proteomic Analysis ◽  
Two Dimensional Gel Electrophoresis ◽  
Maldi Tof Ms ◽  
E Coli ◽  
Comparative Proteomic Analysis ◽  
Maldi Tof ◽  
Northern India ◽  
Flight Mass Spectrometry ◽  
Tof Ms

Background: Most of the proteomic studies in Escherichia coli have focussed on pathogenic strains, while very few studies have studied the commensal strains. It is important to study the commensal strains because under the selective pressure of their habitat, commensal strains might serve as reservoirs of virulent and pathogenic strains. Objective: In this study, we have performed a comparative proteomic analysis of commensal and pathogenic strains of E. coli isolated from a major river flowing through northern India. Methods: Proteins were resolved by two dimensional gel electrophoresis and the differentially expressed proteins were identified using matrix-assisted laser desorption ionization-time of flight mass-spectrometry (MALDI-TOF MS). Results: Many proteins of the commensal strain showed an increased expression compared to the pathogenic strain, of which seventeen proteins were identified by MALDI-TOF MS. Functional classification of these proteins revealed that they belonged to different functional pathways like energy metabolism, nucleotide and nucleoside conversions, translation, biosynthesis of amino acids and motility and energytaxis/chemotaxis. Conclusion: As per the best of our knowledge, this is the first report on comparative proteomic analysis of E. coli commensal and pathogenic strains of aquatic origin. Our results suggest that the increased production of these proteins might play an important role in adaptation of E. coli to a commensal/pathogenic lifestyle. However, further experiments are required to understand the precise role of these proteins in regulating the pathogenicity/commensalism of E. coli.

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