Purification and Characterization of a Secreted Protease from <i>Tetrahymena pyriformis</i>

Banno, Yutaka; Yano, Koh; Nozawa, Yoshinori

doi:10.1111/j.1432-1033.1983.tb07400.x

Cited by 18 publications

(9 citation statements)

References 32 publications

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“…The specific activity for the hydrolysis of HPA at pH 4 compared well with that reported with haemoglobin as substrate (Miiller et al, 1966;Miiller, 1972) whilst the specific activity towards AZC was similar to that reported by others (Suprynowicz & Allewell, 1979;Banno et al, 1982Banno et al, , 1983.…”

Section: Proteolytic Activities Detected and Their Propertiessupporting

confidence: 85%

“…The proteolytic system of the ciliate Tetrahymena pyriformis is probably that most studied amongst protozoan species (Dickie & Liener, 1962;Levy et al, 1976;Banno et al, 1982Banno et al, , 1983. Both acid proteinase and neutral proteinase activities have been described but it is not at all clear how many different types of enzyme are responsible for these.…”

Section: Introductionmentioning

confidence: 99%

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An Analysis of Intracellular Proteolytic Activities of Tetrahymena pyriformis GL

North

Walker

1984

Microbiology

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1977Five different assay conditions were used to analyse the intracellular proteolytic system of Tetrahymena pyriformis GL. Acid proteinase activity was measured using hide powder azure at pH 4 and neutral proteinase activity was measured using hide powder azure and azocasein at pH 8 and two peptide nitroanilides at pH 7. Differences between the activities were apparent from their sensitivity to proteinase inhibitors although all were likely to be due to cysteine proteinases. Indeed all were enhanced by the presence of 1 mM-DTT. The neutral activities could be resolved into at least three forms by ion-exchange chromatography, each form having a different specificity for the substrates used. The acid proteinase activity was similarly separated into different fractions. The intracellular activity of acid proteinase responded differently from that of the neutral activities to culture age or resuspension of cells in fresh medium and starvation buffer. Furthermore, analysis on sucrose density gradients showed that although the neutral activities had a similar subcellular distribution to one another this differed from that of the acid proteinase activity. This suggests that the physiological roles of the acid and neutral proteinases may differ. PAGE using gels containing haemoglobin to detect proteinases was less successful than with other species of protozoa. However, it did reveal multiple forms of acid proteinase.

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Section: Proteolytic Activities Detected and Their Propertiessupporting

confidence: 85%

Section: Introductionmentioning

confidence: 99%

An Analysis of Intracellular Proteolytic Activities of Tetrahymena pyriformis GL

North

Walker

1984

Microbiology

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“…Other ciliate species such as Tetrahymena pyriformis and T. thermophilia have previously been shown to contain and secrete proteases (Banno et al 1983, Straus et al 1992. In these studies, proteases were suggested to be involved in both intracellular and extracellular digestion.…”

Section: Discussionmentioning

confidence: 99%

“…In the scuticociliate parasite of fish Uronema marinum, Lee et al (2003) detected metalloprotease secretion as early as 10 min in ciliate cell suspensions using fluorescence polarisation. They suggested that the metalloproteases are involved in the pathogenesis of this ciliate infection in fishes.Other ciliate species such as Tetrahymena pyriformis and T. thermophilia have previously been shown to contain and secrete proteases (Banno et al 1983, Straus et al 1992. In these studies, proteases were suggested to be involved in both intracellular and extracellular digestion.…”

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confidence: 99%

Identification and partial characterisation of metalloproteases secreted by a Mesanophrys-like ciliate parasite of the Norway lobster Nephrops norvegicus

Small¹,

Neil²,

Taylor³

et al. 2005

Dis. Aquat. Org.

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A ciliate parasite, tentatively identified as Mesanophrys sp. of Norway lobstersNephrops norvegicus, is demonstrated to secrete several proteases into the culture medium (modified Nephrops saline). Analyses using substrate-impregnated sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed 12 activity bands differing greatly in mobility in the gels. The complete inhibition of proteolytic activity by 1,10-phenanthroline indicated that the proteases are of the metallo class. The proteases were active at the physiological temperature (8°C) and haemolymph pH (7.8) of the host. The proteases were selective in the degradation of several host proteins, including the myosin heavy chain, which is a major structural component of lobster muscle. Consequently, these proteases may have important roles in several aspects of the host -parasite interaction including invasion, nutrient uptake by the ciliate, and pathogenesis.KEY WORDS: Ciliate · Parasite · Proteases · Secreted · Pathology Resale or republication not permitted without written consent of the publisherDis Aquat Org 67: [225][226][227][228][229][230][231] 2005 ( Lee et al. 2003); in this case, secreted proteases may be directly responsible for many of the clinical symptoms associated with the disease, such as tissue necrosis.Several reports of Mesanophrys spp. infections in crustaceans have indicated that these ciliates infiltrate and consume host tissues and haemocytes (Poisson 1930, Bang et al. 1972, Messick & Small 1996. The discovery of a Mesanophrys-like parasitic ciliate infecting Norway lobsters Nephrops norvegicus, and isolation of the ciliate via in vitro cultivation, has allowed proteolytic factors that may play a role in the parasite's establishment and proliferation to be investigated. This paper reports on the characterisation of these proteases, and suggests ways in which they may be involved in the pathogenesis of ciliate disease in lobsters. MATERIALS AND METHODSIsolation and cultivation of the ciliate parasite. Routine screening of Nephros norvegicus from the Clyde Sea area for the parasitic dinoflagellate Hematodinium sp. led to the discovery and identification of a ciliate infection in several Norway lobsters (Small et al. 2005). Haemolymph samples were withdrawn aseptically from the base of the fifth pereiopod using a 1 ml disposable syringe and 25-gauge needle following sterilisation of the cuticle with 70% (v/v) ethanol. Ciliate infections were diagnosed by viewing haemolymph smears using light microscopy. Parasitic ciliates were isolated in 3.5 cm well plates with 0.2 ml infected haemolymph added to 5 ml culture medium in each well. The culture medium comprised 10% (v/v) ) and streptomycin (10 µg ml -1 ) were added to inhibit bacterial contamination. The medium was filter-sterilised (0.2 µm) after the addition of all constituents. Cultures were incubated at 8°C. Serial subculturing gave rise to axenic cultures.Experimental subcultures for protease analyses were initiated with the addition of 1 × 10 5 ciliate...

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“…Certain heterotrophic bacteria, including pathogenic bacteria, produce extracellular peptidases (Odagami et al 1994;Simoneni and Palva 1993;Worm and Nybroe 2001). Protozoa also secrete peptidases that enhance degradation and uptake of nitrogen sources (Banno et al 1983). Many different peptidases secreted by these microorganisms act to produce oligopeptides and amino acids, which are essential substrates for microorganism growth and are sources of both organic carbon and nitrogen.…”

Section: Introductionmentioning

confidence: 99%

Seasonal changes in peptidase activities and their properties in the surface water of Lake Shinryu

et al. 2011

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Serial monthly peptidase activities were assayed in the surface water of Lake Shinryu, located in the Chugoku district of Japan, using artificial fluorescent peptidase substrates. The results indicated that the lake water had higher aminopeptidase activities than endopeptidase activities except in November and December, whereas lake water filtrated using membranes with a pore size of 0.2 lm showed higher endopeptidase activities than aminopeptidase activities from June to December, with peaks in June and November. The serial aminopeptidase activity profile was relatively similar to that of the chlorophyll-a concentration. The size distribution of aminopeptidase activities indicated that half of the total activity was retained in the fraction between 5 and 100 lm. These results suggest that phytoplankton participate in the digestion of peptides with aminopeptidase activities. Electrophoretic analysis detected the presence of three peptidase bands (1, 2, and 3) in the lake water that passed through the 0.2-lm membranes. Samples from June, September, and November dominantly contained these active bands in different electrophoretic profiles.

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Purification and Characterization of a Secreted Protease from Tetrahymena pyriformis

Cited by 18 publications

References 32 publications

An Analysis of Intracellular Proteolytic Activities of Tetrahymena pyriformis GL

An Analysis of Intracellular Proteolytic Activities of Tetrahymena pyriformis GL

Identification and partial characterisation of metalloproteases secreted by a Mesanophrys-like ciliate parasite of the Norway lobster Nephrops norvegicus

Seasonal changes in peptidase activities and their properties in the surface water of Lake Shinryu

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