Slicing a protease: Structural features of the ATP‐dependent Lon proteases gleaned from investigations of isolated domains

Rotanova, T. V.; Botos, Istvan; Melnikov, Edward E.; Rasulova, Fatima; Gustchina, Alla; Maurizi, Michael R.; Wlodawer, Alexander

doi:10.1110/ps.052069306

Cited by 85 publications

(70 citation statements)

References 99 publications

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“…Although Spy1536 is annotated in the SF370 genome as a conserved hypothetical protein, it shows similarity to Lon proteases, including a Lon protease PDZ domain, for which reason it was annotated in subsequently published GAS genomes as ATP-dependent endopeptidase Lon. Lon proteases are multidomain enzymes (14,20,52) found in all living organisms and are involved in E. coli in the degradation of naturally unstable and misfolded proteins (52,61). So far, we could not detect any proteolytic activity with the recombinant Spy1536 protein used in these studies, but our data clearly show that the protein is crucial for the surface expression of streptococcal surface proteins and the binding of streptococci to human extracellular matrix proteins.…”

Section: Figmentioning

confidence: 68%

Novel Conserved Group A Streptococcal Proteins Identified by the Antigenome Technology as Vaccine Candidates for a Non-M Protein-Based Vaccine

et al. 2010

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Group A streptococci (GAS) can cause a wide variety of human infections ranging from asymptomatic colonization to life-threatening invasive diseases. Although antibiotic treatment is very effective, when left untreated, Streptococcus pyogenes infections can lead to poststreptococcal sequelae and severe disease causing significant morbidity and mortality worldwide. To aid the development of a non-M protein-based prophylactic vaccine for the prevention of group A streptococcal infections, we identified novel immunogenic proteins using genomic surface display libraries and human serum antibodies from donors exposed to or infected by S. pyogenes. Vaccine candidate antigens were further selected based on animal protection in murine lethal-sepsis models with intranasal or intravenous challenge with two different M serotype strains. The nine protective antigens identified are highly conserved; eight of them show more than 97% sequence identity in 13 published genomes as well as in approximately 50 clinical isolates tested. Since the functions of the selected vaccine candidates are largely unknown, we generated deletion mutants for three of the protective antigens and observed that deletion of the gene encoding Spy1536 drastically reduced binding of GAS cells to host extracellular matrix proteins, due to reduced surface expression of GAS proteins such as Spy0269 and M protein. The protective, highly conserved antigens identified in this study are promising candidates for the development of an M-type-independent, protein-based vaccine to prevent infection by S. pyogenes.

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Section: Figmentioning

confidence: 68%

Novel Conserved Group A Streptococcal Proteins Identified by the Antigenome Technology as Vaccine Candidates for a Non-M Protein-Based Vaccine

et al. 2010

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“…4). Lon proteases belong to the multifunctional family of AAA ϩ proteins (ATPases associated with a variety of cellular activities) that are involved in DNA replication, transcription, membrane fusion, and proteolysis (38). The P. aeruginosa PAO1 genome encodes a Lon protease, PA1803, that is 84% similar to the E. coli Lon protease and one (AsrA/PA0799) that shows 53% similarity to E. coli Lon.…”

Section: Discussionmentioning

confidence: 99%

Involvement of an ATP-Dependent Protease, PA0779/AsrA, in Inducing Heat Shock in Response to Tobramycin in Pseudomonas aeruginosa

Kindrachuk

Fernández

Bains

et al. 2011

Antimicrob Agents Chemother

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The adaptive resistance of Pseudomonas aeruginosa to aminoglycosides is known to occur during chronic lung infections in cystic fibrosis patients in response to nonlethal concentrations of aminoglycosides. Not only is it difficult to achieve high levels of drug throughout the dehydrated mucus in the lung, but also steep oxygen gradients exist across the mucus layer, further reducing the bactericidal activity of aminoglycosides. In this study, microarray analysis was utilized to examine the gene responses of P. aeruginosa to lethal, inhibitory, and subinhibitory concentrations of tobramycin under aerobic and anaerobic conditions. While prolonged exposure to subinhibitory concentrations of tobramycin caused increased levels of expression predominantly of the efflux pump genes mexXY, the greatest increases in gene expression levels in response to lethal concentrations of tobramycin involved a number of heat shock genes and the PA0779 gene (renamed here asrA), encoding an alternate Lon protease. Microarray analysis of an asrA::luxCDABE transposon mutant revealed that the induction of heat shock genes in response to tobramycin in this mutant was significantly decreased compared to that in the parent strain. The level of expression of asrA was induced from an arabinose-inducible promoter to 35-fold greater than wild-type expression levels in the absence of tobramycin, and this overexpression alone caused an increased expression of the heat shock genes, as determined by quantitative PCR (qPCR). This overexpression of asrA conferred short-term protection against lethal levels (4 g/ml) of tobramycin but did not affect the tobramycin MIC. The RpoH heat shock sigma factor was found to be involved in the regulation of asrA in response to both heat shock and tobramycin at the posttranscriptional level. The results of this work suggest that the tobramycin concentration has a significant impact on the gene expression of P. aeruginosa, with lethal concentrations resulting in immediate adaptations conferring short-term protection, such as the induction of the heat shock response, and with subinhibitory concentrations leading to more sustainable long-term protection mechanisms, such as increased efflux.

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“…Each Lon subunit has four domains -the amino-terminal (N) domain that is implicated in the binding of protein substrates, the ATPase (A) domain containing Walker A and B motifs mediating ATP-binding and hydrolysis, the substrate sensor and discriminatory domain (SSD) that discriminates the ATP versus the ADP bound enzyme form, and the carboxyl-terminal (P) domain containing the proteolytic active site (9). In Brevibacillus thermoruber Lon, the N domain may also be involved in subunit oligomerization (10).…”

Section: Structure Of Lonmentioning

confidence: 99%

Functional mechanics of the ATP-dependent Lon protease- lessons from endogenous protein and synthetic peptide substrates

Lee

Suzuki

2008

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

102

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SUMMARYLon, also known as the protease La, is a homo-oligomeric ATP-dependent protease, which is highly conserved in archaea, eubacteria and eukaryotic mitochondria and peroxisomes. Since its discovery, studies have shown that Lon activity is essential for cellular homeostasis, mediating protein quality control and metabolic regulation. This article highlights the discoveries made over the past decade demonstrating that Lon selectively degrades abnormal as well as certain regulatory proteins and thus plays significant roles in maintaining bacterial and mitochondrial function and integrity. In addition, Lon is required in certain pathogenic bacteria, for rendering pathogenicity and host infectivity. Recent research endeavors have been directed towards elucidating the reaction mechanism of the Lon protease by different biochemical and structural biological techniques. In this mini-review, the authors survey the diverse biological roles of Lon, and also place special emphasis on recent discoveries that elucidate the mechanistic aspects of the Lon reaction cycle.

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Slicing a protease: Structural features of the ATP‐dependent Lon proteases gleaned from investigations of isolated domains

Cited by 85 publications

References 99 publications

Novel Conserved Group A Streptococcal Proteins Identified by the Antigenome Technology as Vaccine Candidates for a Non-M Protein-Based Vaccine

Novel Conserved Group A Streptococcal Proteins Identified by the Antigenome Technology as Vaccine Candidates for a Non-M Protein-Based Vaccine

Involvement of an ATP-Dependent Protease, PA0779/AsrA, in Inducing Heat Shock in Response to Tobramycin in Pseudomonas aeruginosa

Functional mechanics of the ATP-dependent Lon protease- lessons from endogenous protein and synthetic peptide substrates

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