Lysosomes in <i>Tetrahymena pyriformis</i> I. Some properties and lysosomal localization of acid hydrolases

Müller, Miklós; Baudhuin, Pierre; Duve, Christian de

doi:10.1002/jcp.1040680211

Cited by 73 publications

(18 citation statements)

References 33 publications

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“…In contrast, MUF-[GlcNAcI3 maximizes productive substrate-enzyme bindings resulting in the cleavage of the glycosidic bond with MUF, and shows simple reaction kinetics (Yang & Hamaguchi 1980). Food vacuole digestive enzymes of protists have been reported to show optimum activity at acid pH (Miiller et al 1966, Nilsson 1979, Nagata & Kirchman 1992. In this study, we found a peak in acid lysozyme activity at pH 4.5 (Fig.…”

Section: Discussionsupporting

confidence: 61%

Digestive enzyme activity as a quantitative measure of protistan grazing: the acid lysozyme assay for bacterivory

Gonzalez¹,

Sherr²,

Sherr³

1993

Mar. Ecol. Prog. Ser.

103

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We propose quantification of the activity of digestive enzymes as a novel way to estimate rates of protist grazing in natural waters. Our first application of this approach was determination of protistan bacterivory by assaying the activity of lysozyme at acid pH. Lysozyme specifically degrades peptidoglycan, a major structural component of prokaryotic cell walls. The basis of the method is determination of lysozyme activity present in protistan food vacuoles by using a fluorochrome-hnked artificial substrate, 4-methylumbelhferyl P-D-N,N'.N"-tnacetylchitotriose as an analogue of peptidoglycan. Measurement of rate of MUF cleavage from the substrate in sonicated samples at acid pH (4.5) distinguishes activity of digestive enzymes present in protistan food vacuoles from extracellular or intracytoplasmic lysozyme activity. Acid lysozyme activity was calibrated against rate of bactenvory estimated using the fluorescently labeled bacteria (FLB) uptake method. Results from the 2 methods were significantly correlated (r2 = 0.98) for both cultures of bacterivorous protists and for estuarine and nearshore seawater samples, over a wide range of rates of bacterivory (lo3 to 106 bacteria ml-' h-') The relation between the 2 variables determined from water samples taken in open North Pacific gyre water had a higher slope compared to that of the other samples. The advantages of the acid lysozyme activity method are that it does not require in vivo incubations, manipulation of live samples. or microscopy, as do other current methods of estimating bacterivory, and that a large number of discrete samples can be quickly processed. Separate calibration of the assay, using alternate measures of bacterivory, is recommended for individual applications.

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Section: Discussionsupporting

confidence: 61%

Digestive enzyme activity as a quantitative measure of protistan grazing: the acid lysozyme assay for bacterivory

Gonzalez¹,

Sherr²,

Sherr³

1993

Mar. Ecol. Prog. Ser.

103

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“…Both enzymes of 0. dc/nicci differed from the induced acid phosphatase of the phytoflagellate Euglena gracilis (Bennun & Blum, 1966), which lost almost all activity within 17 min at 60 "C. Acid phosphatase of the fungus Aspergillus nidulans (Harsanyi & Dorn, 1972) lost about 50 71; activity when heated at 50 "C. The enzymes of 0. danica were active at a wide pH range from 2.2 to 5.2 and the optimum pH for both enzymes was about 4.8. Similar optima for acid phosphatase have been reported for E. gracilis (Blum, 1965), the ciliate Tetrahymena pyriforrnis (Muller, Baudhuin & De Duve, 1966;Lazarus & Scherbaum, 1967), Acantlzanzoeba sp. (Muller, 1969), the green alga Chlorella pyrenoidosa (Knutsen, I 968), the fungus F. monilijiorme (Yoshida & Tamiya,197r), and human erythrocyte acid phosphatase (Fenton & Richardson, 1971) and bone tissue acid phosphatase (Vaes & Jacques, 1965).…”

Section: Discussionsupporting

confidence: 57%

Partial Characterization of the Intra- and Extracellular Acid Phosphatase of an Alga, Ochromonas danica

Patni¹,

Aaronson²

1974

Journal of General Microbiology

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S U M M A R YIntra-and extracellular acid phosphatases were purified 88-and 65-fold respectively from photoheterotrophic Ochroi?zonas danica Pringsheim. The purified enzymes differed in heat inactivation, substrate specificity, and inhibition by several divalent cations and NaF. Intracellular enzyme lost only 30 0; of its activity by heating at 60 "C for 200 min whereas the extracellular enzyme lost 80 ;; . Both enzymes were active over a broad pH range from 2-2 to 5-2 and had an optimum pH of 4-8. Both had broad substrate specificity and differed in their relative ability to hydrolyse /I-glycerophosphate, phenolphthalein diphosphate, glucose-I -phosphate, fructose-I ,6-diphosphate, ADP and ATP. Both were inhibited by Co2+, Zn", Hg", Fe"+, arsenate, tartrate and fluoroacetate but differed in their inhibition by Cu2*, Hg2t and NaF. Intracellular acid phosphatase was more susceptible to inhibition by Hg2-f and NaF, while extracellular acid phosphatase was more susceptible to inhibition by Cu2+. p-Chloromercuribenzoate and urea had no effect on either enzyme's activity. EDTA stimulated the activity of both enzymes. The Kin for the intra-and the extracellular enzymes was 0.5 and 0.33 mM respectively with p-nitrophenyl phosphate as the substrate.

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“…The existence of preexisting lysosomes and their significance for the intracellular degradation of endosomal, heterophagosomal, or autophagosomal contents has already been described for various multicellular (Strauss 1964;Thoenes et al 1970;Pfeiffer 1987) and unicellular eukaryotes (Müller et al 1966;Stockem 1972, 1973). In earlier literature, preexisting lysosomes are frequently denoted as dense bodies, telolysosomes or postlysosomes (Miller and Palade 1964;Gordon et al 1965).…”

Section: Preexisting Lysosomesmentioning

confidence: 91%

Intracellular pathways and degradation of endosomal contents in basal epithelial cells of freshwater sponges (Porifera, Spongillidae)

Hahn-Keser

Stockem

1998

Zoomorphology

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The digestive system expressed by basal epithelial cells of the freshwater sponges Spongilla lacustris and Ephydatia muelleri is mainly represented by a population of 30-50 preexisting lysosomes located in the close vicinity of the central nucleus. The strongly acidic vacuoles (pH 4-4.5) vary in size between 1 and 3 µm, and contain a set of different lysosomal enzymes. Immunocytochemical studies succeeded in the detection of β-hexosaminidase, cathepsin D, acid phosphatase, and α-glucosidase. Endosomes resulting from fluid-phase macropinocytosis, receptor-mediated endocytosis, or phagocytotic activity deliver their exogenous contents to the preexisting lysosomes for enzymatic degradation. Macropinosomes and phagosomes follow a rather reduced intracellular pathway by immediate fusion with the lysosomal compartment, whereas substances conveyed by coated vesicles pass through two additional vacuolar stages, namely early and late endosomes. Early endosomes serve as sorting organelles and segregate various constituents of complex ligands (BSA-AU 6, BSA-AU 12 ) by size into individual late endosomes, which then coalesce with preexisting lysosomes. As a whole, the intracellular pathways and hydrolytic processing of endosomal and phagosomal contents in freshwater sponge cells share certain similarities with the respective mechanisms in cells of higher eukaryotes.& k w d : Abbreviations AU 6/12/18 colloidal gold particles nm · BCECF 2′,7′-bis-(2-carboxyethyl)-5-(6)-carboxyfluorescein · BSA bovine serum albumin · CL-NERF 7′-chlororhodol · CMF-PBS calcium/magnesium-free phosphatebuffered saline · DAMP 3-(2, 4-dinitroanilino)-3′-amino-N-methyldipropylamine · DM-NERF 2′, 7′-dimethylrhodol · DNP dinitrophenol · DTT 1,4-dithiothreitol · EDTA ethylenediaminetetraacetic acid · EGF epidermal growth factor · FCS fetal calf serum · FITC fluorescein-5-isothiocyanate · GTX glycerol-Triton-X 100 · LAMP lysosome-associated membrane protein · LGP lysosomal glycoprotein · LY-CH lucifer yellow carbohydrazide · MPR mannose 6-phosphate receptor · PAGE polyacrylamide gel electrophoresis · PBS phosphate-buffered saline · PEG polyethylene glycol · PFA paraformaldehyde · PIPES 1,4-piperazine-N,N'-bis(2-ethanesulfonic)acid · PMSF phenylmethylsulfonyl fluoride · RT room temperature · SDS sodium dodecyl sulfate · TCA tetrachloric acid · TRIS tris-(hydroxymethyl)aminomethane · TRITC tetramethylrhodamine isothiocyanate& b d y :

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Lysosomes in Tetrahymena pyriformis I. Some properties and lysosomal localization of acid hydrolases

Cited by 73 publications

References 33 publications

Digestive enzyme activity as a quantitative measure of protistan grazing: the acid lysozyme assay for bacterivory

Digestive enzyme activity as a quantitative measure of protistan grazing: the acid lysozyme assay for bacterivory

Partial Characterization of the Intra- and Extracellular Acid Phosphatase of an Alga, Ochromonas danica

Intracellular pathways and degradation of endosomal contents in basal epithelial cells of freshwater sponges (Porifera, Spongillidae)

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