Prevalence of the crpP gene conferring decreased ciprofloxacin susceptibility in enterobacterial clinical isolates from Mexican hospitals

Chávez-Jacobo, Víctor M.; Hernández-Ramírez, Karen C.; Silva-Sánchez, Jesús; Garza–Ramos, Ulises; Barrios-Camacho, Humberto; Ortiz-Alvarado, Rafael; Cervantes, Carlos; Meza-Carmen, Víctor; Ramírez-Díaz, Martha I.

doi:10.1093/jac/dky562

Cited by 28 publications

(28 citation statements)

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“…E. coli JM101 [11] strain was used as the recipient strain for recombinant plasmids [10]. E. coli BL21 (pTrcHisC_crpP) strain was used as the DNA donor of crpP gene as well as a positive control in susceptibility testing and to obtain His-CrpP wild-type (wt) purified protein for enzymatic and infrared assays [8].…”

Section: Bacterial Strains and Culture Mediamentioning

confidence: 99%

“…Homologous crpP genes are distributed across extended spectrum beta-lactamase (ESBL)-producing isolates obtained from Mexican hospitals and confer decreased susceptibility to CIP when transferred to the E. coli J53-3 strain as well as crpP gene from pUM505. All CrpP homologous proteins identified from clinical isolates correspond to small proteins (63-70 amino acids) with an identity of 10.1-43.7 % with respect to the pUM505 CrpP sequence [10]. In general, CrpP homologous proteins are more conserved in the N-terminal region, showing 40-65.7 % identity with respect to the CrpP of pUM505.…”

Section: Introductionmentioning

confidence: 97%

“…In general, CrpP homologous proteins are more conserved in the N-terminal region, showing 40-65.7 % identity with respect to the CrpP of pUM505. However, the C-terminal region displayed low identity (5.7-10.5 %) with the last 30 amino acids of CrpP, suggesting that the N-terminal region could consist of some residues important for CrpP enzymatic activity [10]. In addition, alignment of CrpP homologues and CrpP protein of pUM505 showed that some residues are conserved in all proteins, such as Gly7, Thr8, Asp9, Lys33 and Gly34 (located at the N-terminal region) and Cys40 (located at the C-terminal region), suggesting that these residues could be essential for CrpP function.…”

Section: Introductionmentioning

confidence: 99%

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Identification of Essential Residues for Ciprofloxacin Resistance of Ciprofloxacin-Modifying Enzyme (CrpP) of pUM505

et al. 2020

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The ciprofloxacin-resistance crpP gene, encoded by the pUM505 plasmid, isolated from a P. aeruginosa clinical isolate, confers an enzymatic mechanism of antibiotic phosphorylation, which is ATP-dependent, that decreases ciprofloxacin susceptibility. Homologous crpP genes are distributed across extended spectrum beta-lactamase (ESBL)-producing isolates obtained from Mexican hospitals and which confer decreased susceptibility to CIP. The analysis of sequences of the CrpP of proteins showed that the residues Gly7, Thr8, Asp9, Lys33 and Gly34 (located at the N-terminal region) and Cys40 (located at the C-terminal region) are conserved in all proteins, suggesting that these residues could be essential for CrpP function. The aim of this study was to investigate the amino acids essential to ciprofloxacin resistance, which is conferred by the CrpP protein of pUM505 plasmid. Mutations in the codons encoding Gly7, Asp9, Lys33 and Cys40 of CrpP protein from pUM505 were generated by PCR fusion. The results showed that all mutations generated in CrpP proteins increased ciprofloxacin susceptibility in Escherichia coli . In addition, the CrpP modified proteins were purified and their enzymatic activity on ciprofloxacin was assayed, showing that these modified proteins do not exert catalytic activity on ciprofloxacin. Moreover, by infrared assays it was determined that the modified proteins were are not able to modify the ciprofloxacin molecule. Our findings are the first report that indicate that the amino acids, namely Gly7, Asp9, Lys33 and Cys40, which are conserved in the CrpP proteins, possess an essential role for the enzymatic mechanism that confers ciprofloxacin resistance.

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Section: Bacterial Strains and Culture Mediamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

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Identification of Essential Residues for Ciprofloxacin Resistance of Ciprofloxacin-Modifying Enzyme (CrpP) of pUM505

et al. 2020

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“…Por lo que la visión hacia el futuro es muy preocupante, ya que la resistencia, que este grupo de enzimas proporcionaba y que parecía ser exclusiva para aminoglucósidos, se ha extendido hacia un grupo de antimicrobianos completamente diferente. Además, se ha reportado que CrpP, descubierto inicialmente en un plásmido de P. aeruginosa, se encuentra distribuido en E. coli y en K. pneumoniae (Chávez-Jacobo et al, 2019).…”

Section: Modificación Mediante La Adición De Grupos Funcionalesunclassified

La batalla contra las superbacterias: No más antimicrobianos, no hay ESKAPE

Chávez-Jacobo

2020

TIP RECQB

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La resistencia a los antimicrobianos es uno de los más grandes retos de la medicina moderna. Durante la última década, un grupo de seis bacterias han probado no sólo su capacidad para relativamente “escapar” de los efectos de casi cualquier antimicrobiano, sino también por ser la causa principal de las infecciones hospitalarias. Estos organismos en conjunto se les conoce como ESKAPE, siglas que derivan de la primera letra de la categoría taxonómica género, o sea, del nombre científico de cada una de estas bacterias (Enterococcus spp, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa y Enterobacter spp.). La presente revisión tiene como objetivo describir los principales mecanismos de resistencia asociados a este grupo de bacterias y el impacto que han tenido en el desarrollo de nuevas estrategias antimicrobianas.

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“…[37][38][39] Recently, a novel ciprofloxacin-modifying enzyme gene crpP was detected in pUM505 plasmids, suggesting an increasing trend of PMQR mechanism. 40,41 The actions of Qnr and AAC(6ʹ)-Ib-cr have been systematically investigated. 36,42 Qnr proteins, as pentapeptide repeat protein (PRP) family members, can bind to and protect bacterial DNA gyrase and topoisomerase IV from quinolone inhibition.…”

Section: Introductionmentioning

confidence: 99%

<p>Palmatine Is a Plasmid-Mediated Quinolone Resistance (PMQR) Inhibitor That Restores the Activity of Ciprofloxacin Against QnrS and AAC(6ʹ)-Ib-cr-Producing <em>Escherichia coli</em></p>

Wang

Hao

2020

IDR

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The emergence of plasmid-mediated quinolone resistance (PMQR) is a global challenge in the treatment of clinical disease in both humans and animals and is exacerbated by the presence of different PMQR genes existing in the same bacterial strain. Here, we discovered that a natural isoquinoline alkaloid palmatine extracted from traditional Chinese medicinal plants effectively inhibited the activity of PMQR proteins QnrS and AAC(6′)-Ib-cr. Methods: In total 120 clinical ciprofloxacin-resistant Escherichia coli (E. coli) were screened for the presence of qnrS and aac(6ʹ)-Ib-cr by PCR. Recombinant E. coli that produced QnrS or AAC(6ʹ)-Ib-cr proteins were constructed and the correct expression was confirmed by MALDI/TOF MS analysis and SDS-PAGE. A minimal inhibitory concentration (MICs) assay, growth curve assay and time-kill assay were conducted to evaluate the in vitro antibacterial activity of palmatine and the combination of palmatine and ciprofloxacin. Cytotoxicity assays and mouse thigh infection model were used to evaluate the in vivo synergies. Molecular docking, gyrase supercoiling assay and acetylation assay were used to clarify the mechanism of action. Results: Palmatine effectively restored the activity of ciprofloxacin against qnrS and aac (6ʹ)-Ib-cr-positive E. coli strains in a synergistic manner in vitro. In addition, the combined therapy significantly reduced the bacterial burden in a mouse thigh infection model. Molecular docking revealed that palmatine bound at the functional large loop B of QnrS and Trp102Arg and Asp179Tyr in the binding pocket of AAC(6′)-Ib-cr. Furthermore, interaction analysis confirmed that palmatine reduced the gyrase protective effect of QnrS and the acetylation effect of AAC(6′)-Ib-cr. Conclusion: Our findings suggest that palmatine is a potential efficacious compound to restore PMQR-mediated ciprofloxacin resistance and warrants further preclinical evaluations.

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Prevalence of the crpP gene conferring decreased ciprofloxacin susceptibility in enterobacterial clinical isolates from Mexican hospitals

Cited by 28 publications

References 22 publications

Identification of Essential Residues for Ciprofloxacin Resistance of Ciprofloxacin-Modifying Enzyme (CrpP) of pUM505

Identification of Essential Residues for Ciprofloxacin Resistance of Ciprofloxacin-Modifying Enzyme (CrpP) of pUM505

La batalla contra las superbacterias: No más antimicrobianos, no hay ESKAPE

<p>Palmatine Is a Plasmid-Mediated Quinolone Resistance (PMQR) Inhibitor That Restores the Activity of Ciprofloxacin Against QnrS and AAC(6ʹ)-Ib-cr-Producing <em>Escherichia coli</em></p>

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