scholarly journals Expression, purification, crystallization and preliminary crystallographic analysis of the phosphoglycerate kinase fromAcinetobacter baumannii

2012 ◽  
Vol 68 (7) ◽  
pp. 790-792
Author(s):  
Kayla Baretta ◽  
Craig Garen ◽  
Jiang Yin ◽  
Michael N. G. James
Keyword(s):  
Synchrotron Radiation ◽  
Acinetobacter Baumannii ◽  
Light Source ◽  
Phosphoglycerate Kinase ◽  
Multidrug Resistant ◽  
Crystallographic Analysis ◽  
Lithium Sulfate ◽  
Space Group ◽  
Antibiotic Development ◽  
Potential Target

Acinetobacter baumanniiis a common multidrug-resistant clinical pathogen that is often found in hospitals. TheA. baumanniiphosphoglycerate kinase (AbPGK) is involved in the key energy-producing pathway of glycolysis and presents a potential target for antibiotic development.AbPGK has been expressed and purified; it was crystallized using lithium sulfate as the precipitant. TheAbPGK crystals belonged to space groupP2221. They diffracted to a resolution of 2.5 Å using synchrotron radiation at the Canadian Light Source.

scholarly journals Putative thioredoxin Trx1 fromThermosipho africanusstrain TCF52B: expression, purification and structural determination using S-SAD

2016 ◽  
Vol 72 (6) ◽  
pp. 443-447 ◽  
Author(s):  
Naheda Sahtout ◽  
Jijin R. A. Kuttiyatveetil ◽  
Michel Fodje ◽  
David A. R. Sanders
Keyword(s):  
Synchrotron Radiation ◽  
Light Source ◽  
Dispersion Analysis ◽  
Unit Cell ◽  
Structural Determination ◽  
Biological Processes ◽  
Unit Cell Parameters ◽  
Space Group ◽  
Cell Parameters ◽  
Sequence Identity

Thioredoxin is a small ubiquitous protein that plays a role in many biological processes. A putative thioredoxin, Trx1, fromThermosipho africanusstrain TCF52B, which has low sequence identity to its closest homologues, was successfully cloned, overexpressed and purified. The protein was crystallized using the microbatch-under-oil technique at 289 K in a variety of conditions; crystals grown in 0.2 MMgCl2, 0.1 Mbis-tris pH 6.5, 25%(w/v) PEG 3350, which grew as irregular trapezoids to maximum dimensions of 1.2 × 1.5 × 0.80 mm, were used for sulfur single-wavelength anomalous dispersion analysis. The anomalous sulfur signal could be detected to 2.83 Å resolution using synchrotron radiation on the 08B1-1 beamline at the Canadian Light Source. The crystals belonged to space groupP212121, with unit-cell parametersa= 40.6,b= 41.5,c= 56.4 Å, α = β = γ = 90.0°.

scholarly journals Antimicrobial Activity of Gallium Protoporphyrin IX against Acinetobacter baumannii Strains Displaying Different Antibiotic Resistance Phenotypes

2015 ◽  
Vol 59 (12) ◽  
pp. 7657-7665 ◽  
Author(s):  
Brock A. Arivett ◽  
Steven E. Fiester ◽  
Emily J. Ohneck ◽  
William F. Penwell ◽  
Cynthia M. Kaufman ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Galleria Mellonella ◽  
Therapeutic Option ◽  
Protoporphyrin Ix ◽  
Multidrug Resistant ◽  
Content Type ◽  
Antibiotic Development ◽  
Alveolar Epithelial ◽  
Mueller Hinton ◽  
Novel Therapeutic Strategies

ABSTRACTA paucity of effective, currently available antibiotics and a lull in antibiotic development pose significant challenges for treatment of patients with multidrug-resistant (MDR)Acinetobacter baumanniiinfections. Thus, novel therapeutic strategies must be evaluated to meet the demands of treatment of these often life-threatening infections. Accordingly, we examined the antibiotic activity of gallium protoporphyrin IX (Ga-PPIX) against a collection ofA. baumanniistrains, including nonmilitary and military strains and strains representing different clonal lineages and isolates classified as susceptible or MDR. Susceptibility testing demonstrated that Ga-PPIX inhibits the growth of all tested strains when cultured in cation-adjusted Mueller-Hinton broth, with a MIC of 20 μg/ml. This concentration significantly reduced bacterial viability, while 40 μg/ml killed all cells of theA. baumanniiATCC 19606Tand ACICU MDR isolate after 24-h incubation. Recovery of ATCC 19606Tand ACICU strains from infected A549 human alveolar epithelial monolayers was also decreased when the medium was supplemented with Ga-PPIX, particularly at a 40-μg/ml concentration. Similarly, the coinjection of bacteria with Ga-PPIX increased the survival ofGalleria mellonellalarvae infected with ATCC 19606Tor ACICU. Ga-PPIX was cytotoxic only when monolayers or larvae were exposed to concentrations 16-fold and 1,250-fold higher than those showing antibacterial activity, respectively. These results indicate that Ga-PPIX could be a viable therapeutic option for treatment of recalcitrantA. baumanniiinfections regardless of the resistance phenotype, clone lineage, time and site of isolation of strains causing these infections and their iron uptake phenotypes or the iron content of the media.

scholarly journals Strategies for Rapid Identification of Acinetobacter baumannii Membrane Proteins and Polymyxin B’s Effects

2021 ◽  
Vol 11 ◽  
Author(s):  
Yun Lu ◽  
Xinxin Hu ◽  
Tongying Nie ◽  
Xinyi Yang ◽  
Congran Li ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Sample Preparation ◽  
Acinetobacter Baumannii ◽  
Polymyxin B ◽  
Multidrug Resistant ◽  
Rapid Identification ◽  
Label Free ◽  
Membrane Proteome ◽  
Antibiotic Development ◽  
Digestion Methods

Acinetobacter baumannii, especially multidrug resistant Acinetobacter baumannii, is a notable source of pressure in the areas of public health and antibiotic development. To overcome this problem, attention has been focused on membrane proteins. Different digestion methods and extraction detergents were examined for membrane proteome sample preparation, and label-free quantitative and targeted proteome analyses of the polymyxin B-induced Acinetobacter baumannii ATCC 19606 membrane proteome were performed based on nano LC-MS/MS. Ultracentrifugation of proteins at a speed of 150,000×g, digestion by trypsin, filter-aided sample preparation, and detergents such as lauryldimethylamine-N-oxide were proved as a fast and effective way for identification of membrane proteome by nano LC-MS/MS. Upon treatment with polymyxin B, expression levels of 15 proteins related to membrane structure, transporters, cell surface, and periplasmic space were found to be significantly changed. Furthermore, targeted proteome was also used to confirm these changes. A relatively rapid membrane proteome preparation method was developed, and a more comprehensive view of changes in the Acinetobacter baumannii membrane proteome under polymyxin B pressure was obtained.

scholarly journals Kinetics of 1- and 2-methylallyl + O2 reaction, investigated by photoionisation using synchrotron radiation

2021 ◽  
Author(s):  
Domenik Schleier ◽  
Engelbert Reusch ◽  
Marius Gerlach ◽  
Tobias Preitschopf ◽  
Deb Pratim Mukhopadhyay ◽  
...  
Keyword(s):  
Synchrotron Radiation ◽  
Reaction Kinetics ◽  
Molecular Oxygen ◽  
Light Source ◽  
Storage Ring ◽  
Vacuum Ultraviolet ◽  
Flow Tube ◽  
Tube Reactor ◽  
Swiss Light Source ◽  
Kinetics Of

The reaction kinetics of the isomers of the methylallyl radical with molecular oxygen has been studied in a flow tube reactor at the vacuum ultraviolet (VUV) beamline of the Swiss Light Source storage ring.

scholarly journals Aerosol-based antimicrobial photoinactivation in the lungs: an action spectrum study

2021 ◽  
Author(s):  
Chiara Treghini ◽  
Alfonso Dell’Accio ◽  
Franco Fusi ◽  
Giovanni Romano
Keyword(s):  
Light Source ◽  
A Priori ◽  
Action Spectrum ◽  
Multidrug Resistant ◽  
Theoretical Modelling ◽  
Resistant Bacteria ◽  
Lung Infections ◽  
Killing Action ◽  
Definition Of

AbstractChronic lung infections are among the most diffused human infections, being often associated with multidrug-resistant bacteria. In this framework, the European project “Light4Lungs” aims at synthesizing and testing an inhalable light source to control lung infections by antimicrobial photoinactivation (aPDI), addressing endogenous photosensitizers only (porphyrins) in the representative case of S. aureus and P. aeruginosa. In the search for the best emission characteristics for the aerosolized light source, this work defines and calculates the photo-killing action spectrum for lung aPDI in the exemplary case of cystic fibrosis. This was obtained by applying a semi-theoretical modelling with Monte Carlo simulations, according to previously published methodology related to stomach infections and applied to the infected trachea, bronchi, bronchioles and alveoli. In each of these regions, the two low and high oxygen concentration cases were considered to account for the variability of in vivo conditions, together with the presence of endogenous porphyrins and other relevant absorbers/diffusers inside the illuminated biofilm/mucous layer. Furthermore, an a priori method to obtain the “best illumination wavelengths” was defined, starting from maximizing porphyrin and light absorption at any depth. The obtained action spectrum is peaked at 394 nm and mostly follows porphyrin extinction coefficient behavior. This is confirmed by the results from the best illumination wavelengths, which reinforces the robustness of our approach. These results can offer important indications for the synthesis of the aerosolized light source and definition of its most effective emission spectrum, suggesting a flexible platform to be considered in further applications.

scholarly journals Efficacy of Lysophosphatidylcholine as Direct Treatment in Combination with Colistin against Acinetobacter baumannii in Murine Severe Infections Models

Antibiotics ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 194
Author(s):  
Andrea Miró-Canturri ◽  
Rafael Ayerbe-Algaba ◽  
Manuel Enrique Jiménez-Mejías ◽  
Jerónimo Pachón ◽  
Younes Smani
Keyword(s):  
Acinetobacter Baumannii ◽  
Multidrug Resistant ◽  
Experimental Models ◽  
Antimicrobial Treatment ◽  
Clinical Settings ◽  
Monotherapy Group ◽  
Bacterial Concentration ◽  
Sepsis Model ◽  
Severe Infections ◽  
Stimulation Of

The stimulation of the immune response to prevent the progression of an infection may be an adjuvant to antimicrobial treatment. Here, we aimed to evaluate the efficacy of lysophosphatidylcholine (LPC) treatment in combination with colistin in murine experimental models of severe infections by Acinetobacter baumannii. We used the A. baumannii Ab9 strain, susceptible to colistin and most of the antibiotics used in clinical settings, and the A. baumannii Ab186 strain, susceptible to colistin but presenting a multidrug-resistant (MDR) pattern. The therapeutic efficacies of one and two LPC doses (25 mg/kg/d) and colistin (20 mg/kg/8 h), alone or in combination, were assessed against Ab9 and Ab186 in murine peritoneal sepsis and pneumonia models. One and two LPC doses combined with colistin and colistin monotherapy enhanced Ab9 and Ab186 clearance from spleen, lungs and blood and reduced mice mortality compared with those of the non-treated mice group in both experimental models. Moreover, one and two LPC doses reduced the bacterial concentration in tissues and blood in both models and increased mice survival in the peritoneal sepsis model for both strains compared with those of the colistin monotherapy group. LPC used as an adjuvant of colistin treatment may be helpful to reduce the severity and the resolution of the MDR A. baumannii infection.

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