scholarly journals Dual activity of PNGM-1, a metallo-β-lactamase and tRNase Z, pinpoints the evolutionary origin of subclass B3 metallo-β-lactamases

2019 ◽  
Author(s):  
Jung Hun Lee ◽  
Masayuki Takahashi ◽  
Jeong Ho Jeon ◽  
Lin-Woo Kang ◽  
Mineaki Seki ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Binding Protein ◽  
Clinical Care ◽  
Evolutionary Relationship ◽  
Evolutionary Origin ◽  
Penicillin Binding Protein ◽  
Gram Negative ◽  
Structural Differences ◽  
Trnase Z ◽  
Catalytic Mechanisms

AbstractAntibiotic resistance is a steadily increasing global problem which could lead to a fundamental upheaval in clinical care with the potential to return us to the pre-antibiotic era1-4. The production of β-lactamases, a group of enzymes that confer antibiotic resistance in Gram-negative bacteria, is now one of the major barriers in treating Gram-negative infections5. β-Lactamases are classified according to their catalytic mechanisms into serine β-lactamases and metallo-β-lactamases6,7. There are functional and structural similarities between serine β-lactamases and penicillin-binding proteins, and so serine β-lactamases are thought to have evolved from a penicillin-binding protein7,8. Given the functional and structural differences between serine β-lactamases and metallo-β-lactamases, metallo-β-lactamases are thought to have evolved from a protein other than a penicillin-binding protein, but to date this ancestor remains unknown8-11. We discovered PNGM-1, the first subclass B3 metallo-β-lactamase, in deep-sea sediments that predate the antibiotic era12. Here we discover the dual activity of PNGM-1, pinpointing the evolutionary origin of subclass B3 metallo-β-lactamases. Phylogenetic analysis suggested that PNGM-1 could yield insights into the evolutionary origin of subclass B3 metallo-β-lactamases. We reveal the structural similarities between tRNase Zs and PNGM-1, which prompted us to investigate their evolutionary relationship and the possibility of them possessing dual enzymatic activities. We demonstrate that PNGM-1 has dual activity with both true metallo-β-lactamase and tRNase Z activity, suggesting that PNGM-1 is thought to have evolved from a tRNase Z. We also show kinetic and structural comparisons between PNGM-1 and other proteins including subclass B3 metallo-β-lactamases and tRNase Zs. These comparisons revealed that the B3 metallo-β-lactamase activity of PNGM-1 is a promiscuous activity and subclass B3 metallo-β-lactamases are thought to have evolved through PNGM-1 activity. Our work provides a foundation for the evolution of tRNase Z into subclass B3 metallo-β-lactamases through the dual activity of PNGM-1.

scholarly journals Prevalence, Antibiogram and Molecular Characterization of Comunity-Acquired Methicillin-Resistant Staphylococcus Aureus in AWKA, Anambra Nigeria

2016 ◽  
Vol 10 (1) ◽  
pp. 211-221 ◽  
Author(s):  
Blessing Ike ◽  
Malachy C. Ugwu ◽  
Moses N. Ikegbunam ◽  
David Nwobodo ◽  
Chika Ejikeugwu ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Binding Protein ◽  
Meca Gene ◽  
Latex Agglutination ◽  
Diffusion Method ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Penicillin Binding Protein ◽  
Carriage Rate ◽  
Molecular Features

Objectives:This study evaluated the prevalence, antibiogram and molecular features of CA-MRSA in Awka, Nigeria.Methods:Confirmation of MRSA was done by testing resistance to oxacillin (1µg), cloxacillin (5µg) and cefoxitin(30µg) on sterile Mueller Hinton agar supplemented with 4% sodium chloride. The MRSA strains were subjected to antimicrobial susceptibility testing using Kirby-Bauer disc diffusion method. Minimum inhibitory concentration was determined using agar dilution method. Penicillin binding protein 2a was detected through rapid latex agglutination assay while mecA gene was detected by polymerase chain reaction. A total of 142S. aureusisolates were obtained from 261 samples sourced from Staff, students and fomites of the Faculty of Pharmaceutical SciencesResult:The overall prevalence of MRSA was 22.6%. The carriage rate was higher in females (56.5%) than male (43.5%) and was highest in individuals of 20-30 years of age (57.65%). The MIC of the oxacillin sodium salt ranged from 4-32 μg/ml. The multi-antibiotic resistance indices show that 53.4% had Multiple Antibiotic Resistance Indexing (MARI) higher than 0.2. Penicillin binding protein 2a was detected in 8.4% of MRSA isolates, all from nasal carriage while mecA gene was detected in 5 of isolates.Conclusion:This study showed a very high prevalence of MRSA carriage among studied subjects.

scholarly journals Crystal structures of the transpeptidase domain of the Mycobacterium tuberculosis penicillin-binding protein PonA1 reveal potential mechanisms of antibiotic resistance

FEBS Journal ◽  
2016 ◽  
Vol 283 (12) ◽  
pp. 2206-2218 ◽  
Author(s):  
Ekaterina V. Filippova ◽  
Karen J. Kieser ◽  
Chi-Hao Luan ◽  
Zdzislaw Wawrzak ◽  
Olga Kiryukhina ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Mycobacterium Tuberculosis ◽  
Crystal Structures ◽  
Binding Protein ◽  
Penicillin Binding Protein

Synthetic Peptidoglycan Substrates for Penicillin-Binding Protein 5 of Gram-Negative Bacteria

2004 ◽  
Vol 69 (3) ◽  
pp. 778-784 ◽  
Author(s):  
Dusan Hesek ◽  
Maxim Suvorov ◽  
Ken-ichiro Morio ◽  
Mijoon Lee ◽  
Stephen Brown ◽  
...  
Keyword(s):  
Binding Protein ◽  
Gram Negative Bacteria ◽  
Penicillin Binding Protein ◽  
Gram Negative

scholarly journals On-Chip Isoniazid Exposure of Mycobacterium smegmatis Penicillin-Binding Protein (PBP) Mutant Using Time-Lapse Fluorescent Microscopy

Micromachines ◽  
2018 ◽  
Vol 9 (11) ◽  
pp. 561
Author(s):  
Meltem Elitas
Keyword(s):  
Antibiotic Resistance ◽  
Single Cell ◽  
Mycobacterium Smegmatis ◽  
Single Cell Analysis ◽  
Binding Protein ◽  
Cell Analysis ◽  
Penicillin Binding Protein ◽  
Intact Cells ◽  
The Impact ◽  
Mutant Cells

Antibiotic resistance has been one of the biggest threats to global health. Despite the available prevention and control strategies and efforts in developing new antibiotics, the need remains for effective approaches against antibiotic resistance. Efficient strategies to cope with antimicrobial resistance require a quantitative and deeper understanding of microbial behavior, which can be obtained using different techniques to provide the missing pieces of the current antibiotic-resistance puzzle. Microfluidic-microscopy techniques are among the most promising methods that contribute modernization of traditional assays in microbiology. They provide monitoring and manipulation of cells at micro-scale volumes. Here, we combined population-level, culture-based assays with single-cell resolution, microfluidic-microscopy systems to investigate isoniazid response of Mycobacterium smegmatis penicillin-binding protein (PBP) mutant. This mutant exhibited normal growth in plain medium and sensitivity to stress responses when treated with thermal stress (45 °C), detergent stress (0.1% sodium dodecyl sulfate), acid stress (pH 4.5), and nutrient starvation (1XPBS). The impact of msm0031 transposon insertion on drug-mediated killing was determined for isoniazid (INH, 50 µg/mL), rifampicin (RIF, 200 µg/mL), ethionamide (ETH, 200 µg/mL), and ethambutol (EMB, 5 µg/mL). The PBP mutant demonstrated remarkable isoniazid-killing phenotype in batch culture. Therefore, we hypothesized that single-cell analysis will show increased lysis kinetics and fewer intact cells after drug treatment. However, the single-cell analysis data showed that upon isoniazid exposure, the percentage of the intact PBP mutant cells was 24%, while the percentage of the intact wild-type cells was 4.6%. The PBP mutant cells exhibited decreased cell-lysis profile. Therefore, the traditional culture-based assays were not sufficient to provide insights about the subpopulation of viable but non-culture cells. Consequently, we need more adequate tools to be able to comprehend and fight the antibiotic resistance of bacteria.

scholarly journals A penicillin-binding protein inhibits selection of colistin-resistant, lipooligosaccharide-deficientAcinetobacter baumannii

2016 ◽  
Vol 113 (41) ◽  
pp. E6228-E6237 ◽  
Author(s):  
Joseph M. Boll ◽  
Alexander A. Crofts ◽  
Katharina Peters ◽  
Vincent Cattoir ◽  
Waldemar Vollmer ◽  
...  
Keyword(s):  
Cell Surface ◽  
Outer Membrane ◽  
Lipid A ◽  
Binding Protein ◽  
Resistance Mechanism ◽  
Multidrug Resistant ◽  
Penicillin Binding Protein ◽  
Gram Negative ◽  
Peptidoglycan Cell Wall ◽  
Selection Of

The Gram-negative bacterial outer membrane fortifies the cell against environmental toxins including antibiotics. Unique glycolipids called lipopolysaccharide/lipooligosaccharide (LPS/LOS) are enriched in the cell-surface monolayer of the outer membrane and promote antimicrobial resistance. Colistin, which targets the lipid A domain of LPS/LOS to lyse the cell, is the last-line treatment for multidrug-resistant Gram-negative infections. Lipid A is essential for the survival of most Gram-negative bacteria, but colistin-resistantAcinetobacter baumanniilacking lipid A were isolated after colistin exposure. Previously, strain ATCC 19606 was the onlyA. baumanniistrain demonstrated to subsist without lipid A. Here, we show that otherA. baumanniistrains can also survive without lipid A, but some cannot, affording a unique model to study endotoxin essentiality. We assessed the capacity of 15 clinicalA. baumanniiisolates including 9 recent clinical isolates to develop colistin resistance through inactivation of the lipid A biosynthetic pathway, the products of which assemble the LOS precursor. Our investigation determined that expression of the well-conserved penicillin-binding protein (PBP) 1A, prevented LOS-deficient colony isolation. The glycosyltransferase activity of PBP1A, which aids in the polymerization of the peptidoglycan cell wall, was lethal to LOS-deficientA. baumannii. Global transcriptomic analysis of a PBP1A-deficient mutant and four LOS-deficientA. baumanniistrains showed a concomitant increase in transcription of lipoproteins and their transporters. Examination of the LOS-deficientA. baumanniicell surface demonstrated that specific lipoproteins were overexpressed and decorated the cell surface, potentially compensating for LOS removal. This work expands our knowledge of lipid A essentiality and elucidates a drug resistance mechanism.

scholarly journals Population structure-guided profiling of antibiotic resistance patterns in clinical Listeria monocytogenes isolates from Germany identifies pbpB3 alleles associated with low levels of cephalosporin resistance

2020 ◽  
Author(s):  
Martin A. Fischer ◽  
Sabrina Wamp ◽  
Angelika Fruth ◽  
Franz Allerberger ◽  
Antje Flieger ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Population Structure ◽  
Listeria Monocytogenes ◽  
Allelic Variation ◽  
Binding Protein ◽  
Population Based ◽  
The European Union ◽  
Penicillin Binding Protein ◽  
Minimal Inhibitory Concentrations ◽  
Cephalosporin Resistance

AbstractCase numbers of listeriosis have been increasing in Germany and the European Union during the last decade. In addition reports on the occurrence of antibiotic resistance in Listeria monocytogenes in clinical and environmental isolates are accumulating. The susceptibility towards 14 antibiotics was tested in a selection of clinical L. monocytogenes isolates to get a more precise picture of the development and manifestation of antibiotic resistance in the L. monocytogenes population. Based on the population structure determined by core genome multi locus sequence typing (cgMLST) 544 out of 1,220 sequenced strains collected in Germany between 2009 and 2019 were selected to cover the phylogenetic diversity observed in the clinical L. monocytogenes population. All isolates tested were susceptible towards ampicillin, penicillin and co-trimoxazole - the most relevant antibiotics in the treatment of listeriosis. Resistance to daptomycin and ciprofloxacin was observed in 493 (91%) and in 71 (13%) of 544 isolates, respectively. While all tested strains showed resistance towards ceftriaxone, the minimal inhibitory concentrations (MIC) observed varied widely between 4 mg/L up to >128 mg/L. An allelic variation of the penicillin binding protein gene pbpB3 could be identified as the cause of this difference in ceftriaxone resistance levels. This study is the first population structure-guided analysis of antimicrobial resistance in recent clinical isolates and confirms the importance of penicillin binding protein B3 (PBP B3) for the high level of intrinsic cephalosporin resistance of L. monocytogenes on a population-wide scale.

scholarly journals Cefiderocol Resistance in Acinetobacter baumannii: Roles of β-Lactamases, Siderophore Receptors, and Penicillin Binding Protein 3

2020 ◽  
Vol 64 (11) ◽  
Author(s):  
Saquib Malik ◽  
Monica Kaminski ◽  
David Landman ◽  
John Quale
Keyword(s):  
Acinetobacter Baumannii ◽  
Binding Protein ◽  
Receptor Gene ◽  
Multidrug Resistant ◽  
Resistance Mechanisms ◽  
Penicillin Binding Protein ◽  
Gram Negative ◽  
Content Type ◽  
Siderophore Receptors

ABSTRACT Cefiderocol is a siderophore cephalosporin active against many multidrug-resistant (MDR) Gram-negative pathogens. We examined the resistance mechanisms in 12 Acinetobacter baumannii strains with cefiderocol MICs ranging from ≤0.03 to >32 μg/ml. Cefiderocol resistance could not be explained by β-lactamase activity. Cefiderocol resistance was associated with reduced expression of the siderophore receptor gene pirA. Mutations involving PBP3 may have contributed to resistance in one strain. Additional studies are needed to assess the role of other siderophore receptors.

scholarly journals PBP1a/LpoA but Not PBP1b/LpoB Are Involved in Regulation of the Major β-Lactamase GeneblaAin Shewanella oneidensis

2015 ◽  
Vol 59 (6) ◽  
pp. 3357-3364 ◽  
Author(s):  
Jianhua Yin ◽  
Yiyang Sun ◽  
Yinting Mao ◽  
Miao Jin ◽  
Haichun Gao
Keyword(s):  
Antibiotic Resistance ◽  
Shewanella Oneidensis ◽  
Natural Resistance ◽  
Aquatic Environments ◽  
Gram Negative Bacteria ◽  
Penicillin Binding Protein ◽  
Regulatory Pathway ◽  
Gram Negative ◽  
Content Type ◽  
Class D

ABSTRACTβ-Lactamase production is one of the most important strategies for Gram-negative bacteria to combat β-lactam antibiotics. Studies of the regulation of β-lactamase expression have largely been focused on the class C β-lactamase AmpC, whose induction by β-lactams requires LysR-type regulator AmpR and permease AmpG-dependent peptidoglycan recycling intermediates. InShewanella, which is ubiquitous in aquatic environments and is a reservoir for antibiotic resistance, production of the class D β-lactamase BlaA confers bacteria with natural resistance to many β-lactams. Expression of theblaAgene in the genus representativeShewanella oneidensisis distinct from the AmpC paradigm because of the lack of an AmpR homologue and the presence of an additional AmpG-independent regulatory pathway. In this study, using transposon mutagenesis, we identify proteins that are involved inblaAregulation. Inactivation ofmrcAandlpoA, which encode penicillin binding protein 1a (PBP1a) and its lipoprotein cofactor, LpoA, respectively, drastically enhancesblaAexpression in the absence of β-lactams. Although PBP1b and its cognate, LpoB, also exist inS. oneidensis, their roles inblaAinduction are dispensable. We further show that themrcA-mediatedblaAexpression is independent of AmpG.

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