Identification of Protein Domains on Major Pilin MrkA That Affects the Mechanical Properties of <i>Klebsiella pneumoniae</i> Type 3 Fimbriae

Chan, Chia Han; Chen, Feng Jung; Huang, Ying; Chen, Shin-Yu; Liu, Kuo Liang; Wang, Zhe Chong; Peng, Hwei-Ling; Yew, Tri‐Rung; Liu, Cheng-Hsien; Liou, Gunn‐Guang; Hsu, Ken-Yuh; Chang, Hwan‐You; Hsu, Lih‐Hsing

doi:10.1021/la300224w

Cited by 7 publications

(4 citation statements)

References 41 publications

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“…coli showed the disulfide bond formation between the two cysteines in the similar positions[22]. Furthermore, when we mutated either of the cysteine to alanine we did not detect any expression (data not shown) consistent with a previous study with KP MrkA[20]. In the absence of a crystal structure, our data provide the best indication that MrkA adopts a similar compact structure to FimH with the disulfide bond playing an essential role[22].…”

Section: Resultssupporting

confidence: 88%

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Anti-MrkA Monoclonal Antibodies Reveal Distinct Structural and Antigenic Features of MrkA

Wang¹,

Chen²,

Cvitkovic³

et al. 2017

PLoS ONE

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Antibody therapy against antibiotics resistant Klebsiella pneumoniae infections represents a promising strategy, the success of which depends critically on the ability to identify appropriate antibody targets. Using a target-agnostic strategy, we recently discovered MrkA as a potential antibody target and vaccine antigen. Interestingly, the anti-MrkA monoclonal antibodies isolated through phage display and hybridoma platforms all recognize an overlapping epitope, which opens up important questions including whether monoclonal antibodies targeting different MrkA epitopes can be generated and if they possess different protective profiles. In this study we generated four anti-MrkA antibodies targeting different epitopes through phage library panning against recombinant MrkA protein. These anti-MrkA antibodies elicited strong in vitro and in vivo protections against a multi-drug resistant Klebsiella pneumoniae strain. Furthermore, mutational and epitope analysis suggest that the two cysteine residues may play essential roles in maintaining a MrkA structure that is highly compacted and exposes limited antibody binding/neutralizing epitopes. These results suggest the need for further in-depth understandings of the structure of MrkA, the role of MrkA in the pathogenesis of Klebsiella pneumoniae and the protective mechanism adopted by anti-MrkA antibodies to fully explore the potential of MrkA as an efficient therapeutic target and vaccine antigen.

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Section: Resultssupporting

confidence: 88%

“…An array of 12 peptides covering the full length mature MrkA protein from Klebsiella pneumoniae strain MGH78578 was synthesized(New England Peptide) excluding the signal peptide [20]. Each peptide has 20 amino acid residues, and except for the first and the last peptides each has five amino acid overlapping with flanking peptides.…”

Section: Methodsmentioning

confidence: 99%

Anti-MrkA Monoclonal Antibodies Reveal Distinct Structural and Antigenic Features of MrkA

Wang¹,

Chen²,

Cvitkovic³

et al. 2017

PLoS ONE

View full text Add to dashboard Cite

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“…Bacteria organized in biofilms are difficult to eradicate with common decontamination practices, are generally antimicrobial drugs resistant and could modulate the host immune system [6]. Virulence factors associated with biofilm formation, such as type 1 and type 3 fimbriae, have been implicated in biofilm assembly [7,8,9]. These fimbriae have been identified in K. pneumoniae isolates from the gastrointestinal tract and are associated with pneumonia and urinary tract infection.…”

Section: Introductionmentioning

confidence: 99%

Insights on Klebsiella pneumoniae Biofilms Assembled on Different Surfaces Using Phenotypic and Genotypic Approaches

et al. 2017

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Klebsiella pneumoniae is a prominent etiological agent of healthcare associated infections (HAIs). In this context, multidrug-resistant and biofilm-producing bacteria are of special public health concern due to the difficulties associated with treatment of human infections and eradication from hospital environments. Here, in order to study the impact of medical devices-associated materials on the biofilm dynamics, we performed biofilm phenotypic analyses through a classic and a new scanning electron microscopy (SEM) technique for three multidrug-resistant K. pneumoniae isolates growing on polystyrene and silicone. We also applied whole-genome sequencing (WGS) to search for genetic clues underlying biofilm phenotypic differences. We found major differences in the extracellular polymeric substances (EPS) content among the three strains, which were further corroborated by in-depth EPS composition analysis. WGS analysis revealed a high nucleotide similarity within the core-genome, but relevant differences in the accessory genome that may account for the detected biofilm phenotypic dissimilarities, such as genes already associated with biofilm formation in other pathogenic bacteria (e.g., genes coding haemogglutinins and haemolysins). These data reinforce that the research efforts to defeat bacterial biofilms should take into account that their dynamics may be contingent on the medical devices-associated materials.

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“…MrkA is the major structural component of the fimbrial shaft and MrkD facilitates binding to extracellular-matrix proteins and functions as the fimbrial adhesin. Structural stability of the fimbriae is associated with β strands in the C-terminal region of MrkA (89). Regulation of mrk-gene expression is achieved using a combination of a phosphodiesterase (MrkJ) and a c-di-GMP-binding protein (MrkH) in conjunction with a DNA-binding protein (MrkI).…”

Section: Structure and Genetics Of Fimbriaementioning

confidence: 99%

Epidemiology and Virulence of Klebsiella pneumoniae

2016

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Strains of Klebsiella pneumoniae are frequently opportunistic pathogens implicated in urinary tract and catheter-associated urinary-tract infections of hospitalized patients and compromised individuals. Infections are particularly difficult to treat since most clinical isolates exhibit resistance to several antibiotics leading to treatment failure and the possibility of systemic dissemination. Infections of medical devices such as urinary catheters is a major site of K. pneumoniae infections and has been suggested to involve the formation of biofilms on these surfaces. Over the last decade there has been an increase in research activity designed to investigate the pathogenesis of K. pneumoniae in the urinary tract. These investigations have begun to define the bacterial factors that contribute to growth and biofilm formation. Several virulence factors have been demonstrated to mediate K. pneumoniae infectivity and include, but are most likely not limited to, adherence factors, capsule production, lipopolysaccharide presence, and siderophore activity. The development of both in vitro and in vivo models of infection will lead to further elucidation of the molecular pathogenesis of K. pneumoniae. As for most opportunistic infections, the role of host factors as well as bacterial traits are crucial in determining the outcome of infections. In addition, multidrug-resistant strains of these bacteria have become a serious problem in the treatment of Klebsiella infections and novel strategies to prevent and inhibit bacterial growth need to be developed. Overall, the frequency, significance, and morbidity associated with K. pneumoniae urinary tract infections have increased over many years. The emergence of these bacteria as sources of antibiotic resistance and pathogens of the urinary tract present a challenging problem for the clinician in terms of management and treatment of individuals.

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Identification of Protein Domains on Major Pilin MrkA That Affects the Mechanical Properties of Klebsiella pneumoniae Type 3 Fimbriae

Cited by 7 publications

References 41 publications

Anti-MrkA Monoclonal Antibodies Reveal Distinct Structural and Antigenic Features of MrkA

Anti-MrkA Monoclonal Antibodies Reveal Distinct Structural and Antigenic Features of MrkA

Insights on Klebsiella pneumoniae Biofilms Assembled on Different Surfaces Using Phenotypic and Genotypic Approaches

Epidemiology and Virulence of Klebsiella pneumoniae

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