scholarly journals VIM-15 and VIM-16, Two New VIM-2-Like Metallo-β-Lactamases in Pseudomonas aeruginosa Isolates from Bulgaria and Germany

2008 ◽  
Vol 52 (8) ◽  
pp. 2977-2979 ◽  
Author(s):  
Ines Schneider ◽  
Emma Keuleyan ◽  
Rudolf Rasshofer ◽  
Rumyana Markovska ◽  
Anne Marie Queenan ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Enzyme Activity ◽  
Amino Acid ◽  
Amino Acid Substitution ◽  
Class I ◽  
Class I Integrons

ABSTRACT Two Pseudomonas aeruginosa urine isolates from Bulgaria and Germany produced two new VIM-2 variants. VIM-15 had one amino acid substitution (Tyr218Phe) which caused a significant increase in hydrolytic efficiency. The substitution Ser54Leu, characterizing VIM-16, showed no influence on enzyme activity. Both genes were part of class I integrons located in the chromosome.

Analysis of class I integrons responsible for antibiotics resistance in Pseudomonas aeruginosa

2014 ◽  
Vol 37 (3) ◽  
pp. 241-246
Author(s):  
Man Hwan Oh ◽  
Jangwon Lee ◽  
Seung Kyu Choi ◽  
Seung-Yeol Son
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Class I ◽  
Antibiotics Resistance ◽  
Class I Integrons

scholarly journals Bacterial Two-Hybrid Analysis of Interactions between Region 4 of the ς70 Subunit of RNA Polymerase and the Transcriptional Regulators Rsd from Escherichia coli and AlgQ from Pseudomonas aeruginosa

2001 ◽  
Vol 183 (21) ◽  
pp. 6413-6421 ◽  
Author(s):  
Simon L. Dove ◽  
Ann Hochschild
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Rna Polymerase ◽  
Amino Acid Substitution ◽  
Transcriptional Regulators ◽  
Binding Motif ◽  
E Coli ◽  
Two Hybrid

ABSTRACT A number of transcriptional regulators mediate their effects through direct contact with the ς70 subunit ofEscherichia coli RNA polymerase (RNAP). In particular, several regulators have been shown to contact a C-terminal portion of ς70 that harbors conserved region 4. This region of ς contains a putative helix-turn-helix DNA-binding motif that contacts the −35 element of ς70-dependent promoters directly. Here we report the use of a recently developed bacterial two-hybrid system to study the interaction between the putative anti-ς factor Rsd and the ς70 subunit of E. coli RNAP. Using this system, we found that Rsd can interact with an 86-amino-acid C-terminal fragment of ς70 and also that amino acid substitution R596H, within region 4 of ς70, weakens this interaction. We demonstrated the specificity of this effect by showing that substitution R596H does not weaken the interaction between ς and two other regulators shown previously to contact region 4 of ς70. We also demonstrated that AlgQ, a homolog of Rsd that positively regulates virulence gene expression inPseudomonas aeruginosa, can contact the C-terminal region of the ς70 subunit of RNAP from this organism. We found that amino acid substitution R600H in ς70 fromP. aeruginosa, corresponding to the R596H substitution in E. coli ς70, specifically weakens the interaction between AlgQ and ς70. Taken together, our findings suggest that Rsd and AlgQ contact similar surfaces of RNAP present in region 4 of ς70 and probably regulate gene expression through this contact.

Amino Acid Substitution At Peptide-Binding Pockets of HLA Class I Molecules Adversely Impacts Hematopoietic Cell Transplantation Outcomes

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 467-467
Author(s):  
Joseph Pidala ◽  
Tao Wang ◽  
Michael D Haagenson ◽  
Stephen Spellman ◽  
Medhat Askar ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cell Transplantation ◽  
Amino Acid Substitution ◽  
Acute Gvhd ◽  
Chronic Gvhd ◽  
Peptide Binding ◽  
Multivariable Analysis ◽  
Hla Class I ◽  
Class I ◽  
Increased Risk

Abstract Abstract 467 Background: While donor-recipient disparity at HLA loci is associated with greater risk for severe acute graft vs. host disease (GVHD) and inferior survival after unrelated donor allogeneic hematopoietic cell transplantation (HCT), the impact of amino acid substitution (AAS) at peptide binding pockets of the HLA molecule is incompletely understood. Methods: Adult and pediatric patients who received myeloablative or reduced intensity/non-myeloablative first unrelated bone marrow or peripheral blood stem cell transplantation for AML, ALL, CML or MDS between 1988 and 2009 were included. Donors and patients were fully high resolution matched for HLA-A, B, C, and DRB1 (8/8) or had single mismatch (7/8) at one HLA class I locus. Among 7/8 donor-recipient pairs, cases were categorized based on the presence or absence of the AAS of interest at positions 9, 77, 99, 116, or 156 of the class I molecule. In multivariable analysis accounting for patient, disease, and transplantation variables, we studied the independent impact of AAS at these residues on risk for grade III-IV acute GVHD, chronic GVHD, treatment-related mortality, primary malignancy relapse, and overall survival. We compared 7/8 donor-recipient pairs with AAS of interest to 7/8 pairs without these AAS in the primary analyses. Additionally, we performed this analysis restricted to each HLA class I locus. Results: Donor-recipient pairs were 8/8 matched (n=5282), 7/8 with AAS of interest (n=1713), or 7/8 without AAS of interest (n=318). In multivariable analysis, AAS at position 116 was associated with increased risk for grade III-IV acute GVHD (HR 1.21, 1.04–1.42, p=0.0165). No other significant association was detected between AAS studied and clinical outcomes. In multivariable analysis restricted to each class I HLA locus, we detected the following: Among 7/8 matched pairs with mismatch at HLA-C, AAS at position 116 was associated with increased risk for severe acute GVHD (HR 1.42, 1.13–1.79, p=0.0031) and inferior OS (HR 1.2, 1.01–1.41, p=0.0343). AAS at position 99 was associated with increased TRM (HR 1.37, 1.11–1.69, p=0.0037). Of 7/8 pairs with mismatch at HLA-B, AAS at position 9 was associated with increased chronic GVHD (HR 2.19, 1.31–3.66, p=0.0029). Specific amino acid substitution pairs with frequency > 30 were tested for association with HCT outcomes. None met the significance level of 0.00125, pre-specified for multiple comparisons. Conclusions: These results support the concept that AAS at key peptide-binding residues in the HLA class I molecule are associated with increased risk for severe acute GVHD and lower survival. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Electrogenic Sodium–Sodium Exchange Carried Out by Na,k -Atpase Containing the Amino Acid Substitution Glu779ala

2000 ◽  
Vol 116 (1) ◽  
pp. 61-74 ◽  
Author(s):  
R. Daniel Peluffo ◽  
José M. Argüello ◽  
Jerry B Lingrel ◽  
Joshua R. Berlin
Keyword(s):  
Enzyme Activity ◽  
Amino Acid ◽  
Amino Acid Substitution ◽  
High Capacity ◽  
Outward Current ◽  
Α Subunit ◽  
High Affinity ◽  
High Level ◽  
Kinetics Of ◽  
Insight Into

Na,K -ATPase containing the amino acid substitution glutamate to alanine at position 779 of the α subunit (Glu779Ala) supports a high level of Na-ATPase and electrogenic Na+–Na+ exchange activityin the absence of K +. In microsomal preparations of Glu779Ala enzyme, the Na+ concentration for half maximal activation of Na-ATPase activity was 161 ± 14 mM (n = 3). Furthermore, enzyme activity with 800 mM Na+ was found to be similar in the presence and absence of 20 mM K +. These results showed that Na+, with low affinity, could stimulate enzyme turnover as effectively as K +. To gain further insight into the mechanism of this enzyme activity, HeLa cells expressing Glu779Ala enzyme were voltage clamped with patch electrodes containing 115 mM Na+ during superfusion in K +-free solutions. Electrogenic Na+–Na+ exchange was observed as an ouabain-inhibitable outward current whose amplitude was proportional to extracellular Na+ (Na+o) concentration. At all Na+o concentrations tested (3–148 mM), exchange current was maximal at negative membrane potentials (VM), but decreased as VM became more positive. Analyzing this current at each VM with a Hill equation showed that Na+–Na+ exchange had a high-affinity, low-capacity component with an apparent Na+o affinity at 0 mV (K 00.5) of 13.4 ± 0.6 mM and a low-affinity, high-capacity component with a K 00.5 of 120 ± 13 mM (n = 17). Both high- and low-affinity exchange components were VM dependent, dissipating 30 ± 3% and 82 ± 6% (n = 17) of the membrane dielectric, respectively. The low-affinity, but not the high-affinity exchange component was inhibited with 2 mM free ADP in the patch electrode solution. These results suggest that the high-affinity component of electrogenic Na+–Na+ exchange could be explained by Na+o acting as a low-affinity K + congener; however, the low-affinity component of electrogenic exchange appeared to be due to forward enzyme cycling activated by Na+o binding at a Na+-specific site deep in the membrane dielectric. A pseudo six-state model for the Na,K -ATPase was developed to simulate these data and the results of the accompanying paper (Peluffo, R.D., J.M. Argüello, and J.R. Berlin. 2000. J. Gen. Physiol. 116:47–59). This model showed that alterations in the kinetics of extracellular ion-dependent reactions alone could explain the effects of Glu779Ala substitution on the Na,K -ATPase.

scholarly journals Carbapenem Resistance Mechanisms in Pseudomonas aeruginosa Clinical Isolates

2001 ◽  
Vol 45 (2) ◽  
pp. 480-484 ◽  
Author(s):  
Hyunjoo Pai ◽  
Jong-Won Kim ◽  
Jungmin Kim ◽  
Ji Hyang Lee ◽  
Kang Won Choe ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Amino Acid Substitution ◽  
Carbapenem Resistance ◽  
Resistance Mechanisms ◽  
Clinical Isolates ◽  
Efflux System ◽  
Clinical Strains ◽  
Carbapenem Resistant

ABSTRACT In order to define the contributions of the mechanisms for carbapenem resistance in clinical strains of Pseudomonas aeruginosa, we investigated the presence of OprD, the expressions of the MexAB-OprM and MexEF-OprN systems, and the production of the β-lactamases for 44 clinical strains. All of the carbapenem-resistant isolates showed the loss of or decreased levels of OprD. Three strains overexpressed the MexAB-OprM efflux system by carrying mutations inmexR. These three strains had the amino acid substitution in MexR protein, Arg (CGG) → Gln (CAG), at the position of amino acid 70. None of the isolates, however, expressed the MexEF-OprN efflux system. For the characterization of β-lactamases, at least 13 isolates were the depressed mutants, and 12 strains produced secondary β-lactamases. Based on the above resistance mechanisms, the MICs of carbapenem for the isolates were analyzed. The MICs of carbapenem were mostly determined by the expression of OprD. The MICs of meropenem were two- to four-fold increased for the isolates which overexpressed MexAB-OprM in the background of OprD loss. However, the elevated MICs of meropenem for some individual isolates could not be explained. These findings suggested that other resistance mechanisms would play a role in meropenem resistance in clinical isolates of P. aeruginosa.

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