Microplate fluorescence protease assays test the inhibition of select North American snake venoms' activities with an anti-proteinase library

Price, Joseph A.

doi:10.1016/j.toxicon.2015.06.020

Cited by 7 publications

(6 citation statements)

References 42 publications

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“…Assays were conducted as published [ 29 ] using EnzChek ® Protease Assay, EnzChek ® Elastase Assay, and EnzChek ® Gelatinase/Collagenase Kits from InVitrogen according to kit instructions. The gelatinase substrate has been used for venoms previously [ 31 ].…”

Section: Methodsmentioning

confidence: 99%

“…This is aside from naturally occurring factors in venom-resistant animals, anticipated to often be polypeptides, and thus possibly immunogenic and not therapeutically suitable. Libraries of antiprotease compounds can be screened for their inhibition of protease and collagenase activities using a variety of North American venoms [ 29 ]. Moreover, as others have stated, the possible antivenom mechanisms of action by this plant genus should be explored [ 30 ].…”

Section: Introductionmentioning

confidence: 99%

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An in vitro evaluation of the Native American ethnomedicinal plant Eryngium yuccifolium as a treatment for snakebite envenomation.

Price

2016

J Intercult Ethnopharmacol

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Aim:At least seven North American tribes specifically mention the use of Eryngium (typically roots) as an anti-snake venom therapy. As snake envenomation is an endemic, life-threatening medical risk, is there a scientific basis for the Native American ethnomedicine? Could this be demonstrated in an assay amenable to mechanistic evaluation and high throughput screening for later isolation and possible evaluation as a source for a lead drug?Materials and Methods:Proteases, mainly metalloproteases, are thought to be the main pathological agents in most American snake venoms. Water extracts of four plant parts of Eryngium yuccifolium were tested for enzyme inhibition in three highly sensitive in vitro protease assays, with multiple venoms.Results:Interestingly, activity was found in all plant parts, not just the roots, in the general protease assay, also in the most specific assay for collagenases, but less so for elastases where enzymatic activity was low, and against five species of American snake venoms. Inhibition spared the activity of a mammalian elastase, suggesting it has some specificity. In dose response assays, inhibitory activity in extracts of Eryngium was noticeably more effective than randomly chosen plants and comparable to some others.Conclusions:All data shown here are consistent with pharmacological inhibition of proteases in at least selected venoms of common venomous snakes by Eryngium extracts. Moreover, as the genus is widely distributed in America, the ethnological practice of using this plant as an anti-snake venom treatment is supportable, may have been common, and suggests further bioactivity and phytochemical studies are warranted.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

An in vitro evaluation of the Native American ethnomedicinal plant Eryngium yuccifolium as a treatment for snakebite envenomation.

Price

2016

J Intercult Ethnopharmacol

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“…First, it is widely acknowledged that casein is a suitable model substrate to measure the diversity of proteases in biological systems, as mentioned in the introduction. The hydrolytic action of different protease subtypes on BODIPY FL casein specifically, has been established previously with protease preparations from different origins, such as snake venom (Price, 2015), bacterial and fungal proteases (Ha et al, 2012) and yeast proteases (Hutter et al, 2005). Second, substrate stability is ensured for BODIPY FL based substrates as the BODIPY moiety has low fluorescence bleaching rates and is stable over a pH range from 2 to 11 .…”

Section: Protein Hydrolysis Assay Validationmentioning

confidence: 99%

A high-throughput assay to quantify protein hydrolysis in aerobic and anaerobic wastewater treatment processes

Gaelen

Springael

Smets

2020

Appl Microbiol Biotechnol

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Proteins, an important fraction of the organic matter in wastewater, typically enter a treatment facility as high molecular weight components. These components are degraded by extracellular protein hydrolytic enzymes, denoted as proteases. Adequate protein hydrolysis monitoring is crucial, since protein hydrolysis is often a rate-limiting step in wastewater treatment. However, current monitoring tools lack a high sample throughput and reliable quantification. Here, we present an improved assay for high-throughput protein hydrolysis rate measurements in wastewater treatment applications. A BODIPY FL casein model substrate was implemented in a microplate format for continuous fluorescent quantification. Case studies on a conventional and a high-rate aerobic municipal wastewater treatment plant and a lab-scale, two-stage, anaerobic reactor provided proof-of-concept. The assay presented in this study can help to obtain monitoring-based process insights, which will in turn allow improving biological performance of wastewater treatment installations in the future. Key points-Protein hydrolysis is a crucial step in biological wastewater treatment -Quantification of the protein hydrolysis rate enables in-depth process knowledge -BODIPY FL casein is a suitable model substrate for a protein hydrolysis assay -High sample throughput was obtained with fluorescent hydrolysis quantification

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“…This makes it an inviting candidate for repurposing because of the known safety profile and, thus, potentially reduced development costs [ 8 , 75 , 76 ]. Similarly, MP inhibitors that have previously been developed for cancer treatment such as batimastat, marimastat, and prinomostat could also be considered [ 11 , 12 , 77 , 78 ].…”

Section: Repurposed Drugs and Model Smt Candidates For Enzymatic Imentioning

confidence: 99%

Developing Small Molecule Therapeutics for the Initial and Adjunctive Treatment of Snakebite

Bulfone

Samuel

Bickler

et al. 2018

Journal of Tropical Medicine

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The World Health Organization (WHO) recently added snakebite envenoming to the priority list of Neglected Tropical Diseases (NTD). It is thought that ~75% of mortality following snakebite occurs outside the hospital setting, making the temporal gap between a bite and antivenom administration a major therapeutic challenge. Small molecule therapeutics (SMTs) have been proposed as potential prereferral treatments for snakebite to help address this gap. Herein, we discuss the characteristics, potential uses, and development of SMTs as potential treatments for snakebite envenomation. We focus on SMTs that are secretory phospholipase A2 (sPLA2) inhibitors with brief exploration of other potential drug targets on venom molecules.

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Microplate fluorescence protease assays test the inhibition of select North American snake venoms' activities with an anti-proteinase library

Cited by 7 publications

References 42 publications

An in vitro evaluation of the Native American ethnomedicinal plant Eryngium yuccifolium as a treatment for snakebite envenomation.

An in vitro evaluation of the Native American ethnomedicinal plant Eryngium yuccifolium as a treatment for snakebite envenomation.

A high-throughput assay to quantify protein hydrolysis in aerobic and anaerobic wastewater treatment processes

Developing Small Molecule Therapeutics for the Initial and Adjunctive Treatment of Snakebite

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