Enzymatic degradation of a prion-like protein, Sup35NM-His6

Wang, Jeng-Jie; Borwornpinyo, Rattana; Odetallah, Nasser; Shih, Jason C. H.

doi:10.1016/j.enzmictec.2004.12.023

Cited by 14 publications

(7 citation statements)

References 30 publications

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“…Furthermore, recent work from Barlow et al38 on adult cement has shown around 50% of the adhesive to be made up of amyloid‐like β‐sheet proteins. Subtilisin A is highly effective in breaking down amyloid39, 40 and while the available evidence suggests that the cyprid permanent cement and adult cement differ,9, 18, 41 the pronounced effect of Subtilisin A on cyprid permanent adhesion may point to amyloid content in cyprid cement.…”

Section: Discussionmentioning

confidence: 99%

Active Enzyme Nanocoatings Affect Settlement of Balanus amphitrite Barnacle Cyprids

Tasso

Conlan

Clare

et al. 2011

Adv Funct Materials

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Balanus amphitrite cyprids produce complex adhesive substances that enable their attachment to surfaces and impart a strong detachment resistance from most immersed substrata. The colonization of man‐made structures by barnacle cyprids and other marine organisms is a troublesome and costly phenomenon, for which controlling strategies are actively sought. In this work, we expand previous investigations about the susceptibility of cyprid adhesives to unpurified proteases in solution by evaluating the interplay between these secreted biomolecules and a surface‐confined purified protease. The strategy involved the covalent immobilization of the enzyme Subtilisin A to maleic anhydride copolymer thin films through the spontaneous reaction of anhydride moieties with lysine side chains. This enabled the production of bioactive layers of tunable enzyme surface concentration and activity, which were utilized to systematically evaluate the effect of the immobilized enzyme on cyprid settlement and exploratory behavior. Surfaces of increasing enzyme activity displayed a gradual decrease in cyprid settlement levels (approaching inhibition) as well as an increase in post‐settlement adhesion failure (evidenced by significant numbers of detached metamorphosed individuals). High activities of the bound enzyme also affected pre‐settlement behavior of cyprids, reducing the velocity and total distance moved while increasing the amount and speed of meander compared to the controls. The here‐reported low enzyme surface concentrations found to be remarkably effective at reducing cyprid settlement hold promise for the use of immobilized enzymes in the control of marine biofouling.

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Section: Discussionmentioning

confidence: 99%

Active Enzyme Nanocoatings Affect Settlement of Balanus amphitrite Barnacle Cyprids

Tasso

Conlan

Clare

et al. 2011

Adv Funct Materials

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“…Being nonpathogenic and fairly easy to prepare, Sup35NM is a good candidate as a prion model for many research and technological applications. A similar protein molecule was recently developed and tested for its prion‐like properties (Wang et al 2005). Sup35NM and this molecule will be compared to determine the abilities of each to serve as a safe prion marker for disinfection processes and metabolic studies.…”

Section: Discussionmentioning

confidence: 99%

Characterization and enzymatic degradation of Sup35NM, a yeast prion‐like protein

et al. 2005

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Transmissible spongiform encephalopathies (TSEs) are believed to be caused by an unconventional infectious agent, the prion protein. The pathogenic and infectious form of prion protein, PrP Sc , is able to aggregate and form amyloid fibrils, very stable and resistant to most disinfecting processes and common proteases. Under specific conditions, PrP Sc in bovine spongiform encephalopathy (BSE) brain tissue was found degradable by a bacterial keratinase and some other proteases. Since this disease-causing prion is infectious and dangerous to work with, a model or surrogate protein that is safe is needed for the in vitro degradation study. Here a nonpathogenic yeast prion-like protein, Sup35NM, cloned and overexpressed in E. coli, was purified and characterized for this purpose. Aggregation and deaggregation of Sup35NM were examined by electron microscopy, gel electrophoresis, Congo red binding, fluorescence, and Western blotting. The degradation of Sup35NM aggregates by keratinase and proteinase K under various conditions was studied and compared. These results will be of value in understanding the mechanism and optimization of the degradation process.Keywords: BSE; prion; PrP Sc ; Sup35NM; yeast prion; prion surrogate protein; enzymatic degradation Prion diseases, or transmissible spongiform encephalopathies (TSEs), are a unique group of fatal neurodegenerative disorders found in humans and animals (Prusiner et al. 1998). They include diseases such as kuru and Creutzfeld-Jakob disease (CJD) in humans, scrapie in sheep, bovine spongiform encephalopathy (BSE, widely known as mad cow disease) in cattle (Parchi and Gambetti 1995), and TSEs in other mammals (Burger and Hartsough 1965;Williams and Young 1980;Leggett et al. 1990). All of these diseases are characterized by extensive neuronal loss giving brain tissue a spongy appearance.The pathological isoform of prion protein (PrP Sc ) aggregates in diseased tissues and, in common conditions, is resistant to digestion by proteases (Caughey et al. 1991;Pan et al. 1993). It was recently found that PrP Sc in diseased tissues can be degraded by a feather-degrading keratinase and other microbial proteases when the tissue homogenate was preheated at 115 C (Langeveld et al. 2003). Independently, mouse bioassays showed a significant reduction of infectivity after a proteolytic inactivation process (McLeod et al. 2004). These results indicated a potential method of enzymatic inactivation of prion protein.PrP Sc is pathogenic and dangerous to handle. A mutant recombinant prion produced in Escherichia coli without glycosylation was recently found to be able to induce neurologic dysfunction (Legname et al. 2004). Experimentation with prion tissues must be strictly controlled and is inordinately expensive. A nonpathogenic prion surrogate protein which has similar physical-chemical properties Reprint requests to: Ching-Ying Chen, Department of Poultry Science, North Carolina State University, Raleigh, NC 27695-7608, USA; e-mail: cchen8@ncsu.edu; fax: (919) 515-2625.Article ...

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“…This is due to the possible development of antifungals that reduce the level of biofilm formation and the potential use of enzyme-containing formulations for the treatment of neurodegenerative diseases in humans. Information about such proteases hydrolyzing amyloid proteins is presented in Table 4 [114][115][116][117][118][119][120][121][122][123][124][125][126][127][128][129].…”

Section: Enzymes Hydrolyzing Fungal Proteins With Amyloid Characteris...mentioning

confidence: 99%

Metal Nanomaterials and Hydrolytic Enzyme-Based Formulations for Improved Antifungal Activity

Lyagin

Aslanli

Domnin

et al. 2023

IJMS

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Active research of metal-containing compounds and enzymes as effective antifungal agents is currently being conducted due to the growing antifungal resistance problem. Metals are attracting special attention due to the wide variety of ligands that can be used for them, including chemically synthesized and naturally obtained variants as a result of the so-called “green synthesis”. The main mechanism of the antifungal action of metals is the triggering of the generation and accumulation of reactive oxygen species (ROS). Further action of ROS on various biomolecules is nonspecific. Various hydrolytic enzymes (glucanases and proteases), in turn, exhibit antifungal properties by affecting the structural elements of fungal cells (cell walls, membranes), fungal quorum sensing molecules, fungal own protective agents (mycotoxins and antibiotics), and proteins responsible for the adhesion and formation of stable, highly concentrated populations in the form of biofilms. A wide substrate range of enzymes allows the use of various mechanisms of their antifungal actions. In this review, we discuss the prospects of combining two different types of antifungal agents (metals and enzymes) against mycelial fungi and yeast cells. Special attention is paid to the possible influence of metals on the activity of the enzymes and the possible effects of proteins on the antifungal activity of metal-containing compounds.

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Enzymatic degradation of a prion-like protein, Sup35NM-His6

Cited by 14 publications

References 30 publications

Active Enzyme Nanocoatings Affect Settlement of Balanus amphitrite Barnacle Cyprids

Active Enzyme Nanocoatings Affect Settlement of Balanus amphitrite Barnacle Cyprids

Characterization and enzymatic degradation of Sup35NM, a yeast prion‐like protein

Metal Nanomaterials and Hydrolytic Enzyme-Based Formulations for Improved Antifungal Activity

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