scholarly journals Enzymatic and structural characterization of the Naegleria fowleri glucokinase

2018 ◽  
Author(s):  
Jillian E. Milanes ◽  
Jimmy Suryadi ◽  
Jan Abendroth ◽  
Wesley C. Van Voorhis ◽  
Kayleigh F. Barrett ◽  
...  
Keyword(s):  
Treatment Options ◽  
Structural Similarity ◽  
Amino Acid Identity ◽  
Pentose Phosphate ◽  
Mammalian Host ◽  
Binding Modes ◽  
Naegleria Fowleri ◽  
Therapeutic Development ◽  
Small Collection ◽  
Nægleria Fowleri

ABSTRACTInfection with the free-living amoeba Naegleria fowleri leads to life-threatening primary amoebic meningoencephalitis. Efficacious treatment options for these infections are limited and the mortality rate is very high (~98%). Parasite metabolism may provide suitable targets for therapeutic design. Like most other organisms, glucose metabolism is critical for parasite viability, being required for growth in culture. The genome of the parasite encodes a single glucose phosphorylating enzyme, a glucokinase (Glck). The products of this enzyme are required for both glycolysis and the pentose phosphate pathway. The N. fowleri Glck (NfGlck) shares limited (25%) amino acid identity with the mammalian host enzyme (HsGlck), suggesting that parasite-specific inhibitors with anti-amoeba activity could be generated. Following heterologous expression, NfGlck was found to have a limited hexose substrate range, with greatest activity observed with glucose. The enzyme had apparent Km values of 42.5 ± 7.3 μM and 141.6 ± 9.9 μM for glucose and ATP, respectively. The NfGlck structure was determined and refined to 2.2 Å resolution, revealing that the enzyme shares greatest structural similarity with the Trypanosoma cruzi Glck. These similarities include binding modes and binding environments for substrates. To identify inhibitors of NfGlck, we screened a small collection of inhibitors of glucose phosphorylating enzymes and identified several small molecules with IC50 values < 1 μM that may prove useful as hit chemotypes for further lead and therapeutic development against N. fowleri.

scholarly journals Enzymatic and Structural Characterization of the Naegleria fowleri Glucokinase

2019 ◽  
Vol 63 (5) ◽  
Author(s):  
Jillian E. Milanes ◽  
Jimmy Suryadi ◽  
Jan Abendroth ◽  
Wesley C. Van Voorhis ◽  
Kayleigh F. Barrett ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
Homo Sapiens ◽  
Treatment Options ◽  
Amino Acid Identity ◽  
Pentose Phosphate ◽  
Mammalian Host ◽  
Binding Modes ◽  
Naegleria Fowleri ◽  
Content Type ◽  
Nægleria Fowleri

ABSTRACT Infection with the free-living amoeba Naegleria fowleri leads to life-threatening primary amoebic meningoencephalitis. Efficacious treatment options for these infections are limited, and the mortality rate is very high (∼98%). Parasite metabolism may provide suitable targets for therapeutic design. Like most other organisms, glucose metabolism is critical for parasite viability, being required for growth in culture. The first enzyme required for glucose metabolism is typically a hexokinase (HK), which transfers a phosphate from ATP to glucose. The products of this enzyme are required for both glycolysis and the pentose phosphate pathway. However, the N. fowleri genome lacks an obvious HK homolog and instead harbors a glucokinase (Glck). The N. fowleri Glck (NfGlck) shares limited (25%) amino acid identity with the mammalian host enzyme (Homo sapiens Glck), suggesting that parasite-specific inhibitors with anti-amoeba activity can be generated. Following heterologous expression, NfGlck was found to have a limited hexose substrate range, with the greatest activity observed with glucose. The enzyme had apparent Km values of 42.5 ± 7.3 μM and 141.6 ± 9.9 μM for glucose and ATP, respectively. The NfGlck structure was determined and refined to 2.2-Å resolution, revealing that the enzyme shares greatest structural similarity with the Trypanosoma cruzi Glck. These similarities include binding modes and binding environments for substrates. To identify inhibitors of NfGlck, we screened a small collection of inhibitors of glucose-phosphorylating enzymes and identified several small molecules with 50% inhibitory concentration values of <1 μM that may prove useful as hit chemotypes for further leads and therapeutic development against N. fowleri.

scholarly journals Naegleria fowleri: Pathogenesis, Diagnosis, and Treatment Options

2015 ◽  
Vol 59 (11) ◽  
pp. 6677-6681 ◽  
Author(s):  
Eddie Grace ◽  
Scott Asbill ◽  
Kris Virga
Keyword(s):  
Case Reports ◽  
Treatment Options ◽  
Clinical Manifestations ◽  
Naegleria Fowleri ◽  
Content Type ◽  
High Profile ◽  
Free Living Amoeba ◽  
Nægleria Fowleri ◽  
Amoebic Meningoencephalitis

ABSTRACTNaegleria fowlerihas generated tremendous media attention over the last 5 years due to several high-profile cases. Several of these cases were followed very closely by the general public.N. fowleriis a eukaryotic, free-living amoeba belonging to the phylum Percolozoa.Naegleriaamoebae are ubiquitous in the environment, being found in soil and bodies of freshwater, and feed on bacteria found in those locations. WhileN. fowleriinfection appears to be quite rare compared to other diseases, the clinical manifestations of primary amoebic meningoencephalitis are devastating and nearly always fatal. Due to the rarity ofN. fowleriinfections in humans, there are no clinical trials to date that assess the efficacy of one treatment regimen over another. Most of the information regarding medication efficacy is based on either case reports orin vitrostudies. This review will discuss the pathogenesis, diagnosis, pharmacotherapy, and prevention ofN. fowleriinfections in humans, including a brief review of all survivor cases in North America.

Naegleria fowleri: diagnosis, treatment options and pathogenesis

2019 ◽  
Vol 7 (2) ◽  
pp. 67-80 ◽  
Author(s):  
Mohammad Ridwane Mungroo ◽  
Naveed Ahmed Khan ◽  
Ruqaiyyah Siddiqui
Keyword(s):  
Treatment Options ◽  
Naegleria Fowleri ◽  
Nægleria Fowleri

scholarly journals Laurinterol from Laurencia johnstonii eliminates Naegleria fowleri triggering PCD by inhibition of ATPases

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Iñigo Arberas-Jiménez ◽  
Sara García-Davis ◽  
Aitor Rizo-Liendo ◽  
Ines Sifaoui ◽  
María Reyes-Batlle ◽  
...  
Keyword(s):  
Cell Death ◽  
Apoptotic Cell ◽  
Treatment Options ◽  
Opportunistic Pathogen ◽  
Current Treatment ◽  
Ion Pump ◽  
Naegleria Fowleri ◽  
Trophozoite Stage ◽  
Nægleria Fowleri

Abstract Primary amoebic encephalitis (PAM) is a lethal disease caused by the opportunistic pathogen, Naegleria fowleri. This amoebic species is able to live freely in warm aquatic habitats and to infect children and young adults when they perform risk activities in these water bodies such as swimming or splashing. Besides the need to increase awareness of PAM which will allow an early diagnosis, the development of fully effective therapeutic agents is needed. Current treatment options are amphotericin B and miltefosine which are not fully effective and also present toxicity issues. In this study, the in vitro activity of various sesquiterpenes isolated from the red alga Laurencia johnstonii were tested against the trophozoite stage of a strain of Naegleria fowleri. Moreover, the induced effects (apoptotic cell death) of the most active compound, laurinterol (1), was evaluated by measuring DNA condensation, damages at the mitochondrial level, cell membrane disruption and production of reactive oxygen species (ROS). The obtained results demonstrated that laurinterol was able to eliminate the amoebae at concentrations of 13.42 ± 2.57 µM and also to induced programmed cell death (PCD) in the treated amoebae. Moreover, since ATP levels were highly affected and laurinterol has been previously reported as an inhibitor of the Na+/K+-ATPase sodium–potassium ion pump, comparison with known inhibitors of ATPases were carried out. Our results points out that laurinterol was able to inhibit ENA ATPase pump at concentrations 100 times lower than furosemide.

Efficacy of a combined system of copper and silver and free chlorine for inactivation of Naegleria fowleri amoebas in water

1995 ◽  
Vol 31 (5-6) ◽  
pp. 119-122 ◽  
Author(s):  
J. M. Cassells ◽  
M. T. Yahya ◽  
C. P. Gerba ◽  
J. B. Rose
Keyword(s):  
Synergistic Effect ◽  
Exposure Time ◽  
Silver Ions ◽  
Free Chlorine ◽  
Log10 Reduction ◽  
Naegleria Fowleri ◽  
Combined System ◽  
Water Ph ◽  
Nægleria Fowleri ◽  
Time Required

Electrolytically generated copper and silver ions (400:40 and 800:80 μg/l) were evaluated, separately and combined with 1.0 mg/l free chlorine, for their efficacy in reducing the viable numbers of Naegleria fowleri amoebas in water (pH 7.3 and 23-25°C). Inactivation rates (k = log10 reduction/min) and T99 values (exposure time required to achieve a 99% or a 2 log10 reduction) of the disinfectants were determined. Copper and silver alone, at ratio of 400:40 to 800:80 μg/l caused no significant inactivation of N. fowleri even after 72 hours of exposure (k = 0.00017 and 0.00013, respectively). Addition of 1.0 mg/l free chlorine to water which contained 400:40 or 800:80 μg/l copper and silver resulted in enhanced inactivation rates (k = 0.458 and 0.515, respectively) compared to either chlorine alone (k = 0.33) or the metals alone. Water containing 800:80 μg/l copper and silver with 1.0 mg/l chlorine showed a T99 value of 3.9 minutes, while chlorine alone showed a T99 of 6.1 minutes. Enhanced inactivation of N. fowleri by a combined system of free chlorine and copper and silver may be attributed to the different mechanism that each disinfectant utilizes in inactivating the amoebas, and may suggest a synergistic effect.

Biological and In silico Studies on Synthetic Analogues of Tyrosine Betaine as Inhibitors of Neprilysin - A Drug Target for the Treatment of Heart Failure

2018 ◽  
Vol 24 (17) ◽  
pp. 1899-1904
Author(s):  
Daniel Fabio Kawano ◽  
Marcelo Rodrigues de Carvalho ◽  
Mauricio Ferreira Marcondes Machado ◽  
Adriana Karaoglanovic Carmona ◽  
Gilberto Ubida Leite Braga ◽  
...  
Keyword(s):  
Heart Failure ◽  
Secondary Metabolites ◽  
Structural Similarity ◽  
Structural Features ◽  
Major Constituent ◽  
Binding Modes ◽  
Starting Point ◽  
In Silico Studies ◽  
Nep Inhibition

Background: Fungal secondary metabolites are important sources for the discovery of new pharmaceuticals, as exemplified by penicillin, lovastatin and cyclosporine. Searching for secondary metabolites of the fungi Metarhizium spp., we previously identified tyrosine betaine as a major constituent. Methods: Because of the structural similarity with other inhibitors of neprilysin (NEP), an enzyme explored for the treatment of heart failure, we devised the synthesis of tyrosine betaine and three analogues to be subjected to in vitro NEP inhibition assays and to molecular modeling studies. Results: In spite of the similar binding modes with other NEP inhibitors, these compounds only displayed moderate inhibitory activities (IC50 ranging from 170.0 to 52.9 µM). However, they enclose structural features required to hinder passive blood brain barrier permeation (BBB). Conclusions: Tyrosine betaine remains as a starting point for the development of NEP inhibitors because of the low probability of BBB permeation and, consequently, of NEP inhibition at the Central Nervous System, which is associated to an increment in the Aβ levels and, accordingly, with a higher risk for the onset of Alzheimer's disease.

scholarly journals A Novel Cysteine Protease Inhibitor of Naegleria fowleri That Is Specifically Expressed during Encystation and at Mature Cysts

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 388
Author(s):  
Hương Giang Lê ◽  
A-Jeong Ham ◽  
Jung-Mi Kang ◽  
Tuấn Cường Võ ◽  
Haung Naw ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Cysteine Protease ◽  
Expression Profiles ◽  
Critical Role ◽  
Cyst Formation ◽  
Cysteine Proteases ◽  
Cysteine Protease Inhibitor ◽  
Naegleria Fowleri ◽  
Nægleria Fowleri ◽  
Cystatin Family

Naegleria fowleri is a free-living amoeba that is ubiquitous in diverse natural environments. It causes a fatal brain infection in humans known as primary amoebic meningoencephalitis. Despite the medical importance of the parasitic disease, there is a great lack of knowledge about the biology and pathogenicity of N. fowleri. In this study, we identified and characterized a novel cysteine protease inhibitor of N. fowleri (NfCPI). NfCPI is a typical cysteine protease inhibitor belonging to the cystatin family with a Gln-Val-Val-Ala-Gly (QVVAG) motif, a characteristic motif conserved in the cystatin family of proteins. Bacterially expressed recombinant NfCPI has a dimeric structure and exhibits inhibitory activity against several cysteine proteases including cathespin Bs of N. fowleri at a broad range of pH values. Expression profiles of nfcpi revealed that the gene was highly expressed during encystation and cyst of the amoeba. Western blot and immunofluorescence assays also support its high level of expression in cysts. These findings collectively suggest that NfCPI may play a critical role in encystation or cyst formation of N. fowleri by regulating cysteine proteases that may mediate encystation or mature cyst formation of the amoeba. More comprehensive studies to investigate the roles of NfCPI in encystation and its target proteases are necessary to elucidate the regulatory mechanism and the biological significance of NfCPI.

Transmission of Naegleria fowleri between Mice

1983 ◽  
Vol 69 (1) ◽  
pp. 249 ◽  
Author(s):  
R. G. May ◽  
D. T. John
Keyword(s):  
Naegleria Fowleri ◽  
Nægleria Fowleri
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