Tissue Fraction-Past and Present

Duve, Christian de

doi:10.1083/jcb.50.1.20d

Cited by 368 publications

(148 citation statements)

References 97 publications

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“…After digitonin treatment the membranes related to this group undergo a very typical modification of their electron microscope appearance. An obvious relationship with isolated plasma membranes is thus established, since the latter suffer the same modifications of their density distribution (1,12) and of their morphology (12,34) on treatment with (Table II of reference 7).…”

Section: Resultsmentioning

confidence: 86%

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Analytical Study of Microsomes and Isolated Subcellular Membranes From Rat Liver

Amar-Costesec

Wibo

Thinessempoux

et al. 1974

The Journal of Cell Biology

215

112

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lsopycnic equilibration and sedimentation rate studies of rat liver microsomes led previously to the assignment of microsomal constituents into group al (monoamine oxidase), group a2 (5'-nucleotidase, alkaline phosphodiesterase I, alkaline phosphatase and cholesterol), group a3 (galactosyltransferase), group b (NADH cytochrome ¢ reductase, NADPH cytochrome c reductase, aminopyrine demethylase, cytochrome b5 and P 450), and group c (glucose 6-phosphatase, esterase, nucleoside diphosphatase, 13-glucuronidase and glucuronyltransferase).Confirmation and extension of the assignment into groups has been obtained by studying the differential effect of the reagents digitonin, EDTA, and PPi. Digitonin specifically affected the equilibrium density only of the group a2 and (to a lesser extent) group a3, and not of groups b and c under conditions which preserved the structure-linked latency of nucleoside diphosphatase and galactosyltransferase. Within experimental error the rate of sedimentation of all microsomal constituents was unaffected. The morphological appearance under the electron microscope was indistinguishable from that of nondigitonin-treated microsomes, except that a few smooth membranes (< 10%) exhibited broken-looking profiles.Treatment of microsomes with EDTA or PPi detached a substantial part of RNA and released protein in excess over the amount accountable for by detachment of ribosome constituents. This detachment was confirmed by electron microscopy. EDTA and PPi decreased markedly the equilibrium density and the density dispersion of groups b and c, due mainly to the uncoating of rough

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Section: Resultsmentioning

confidence: 86%

“…The results of dig±ton±n, PPifl and E D T A t r e a t m e n t are reported here. Preliminary accounts of this work have already appeared (1,2,5,12,28,29,34).…”

Section: Dase (Group Al) and Galactosyltransferase (Group A3)mentioning

confidence: 93%

Analytical Study of Microsomes and Isolated Subcellular Membranes From Rat Liver

Amar-Costesec

Wibo

Thinessempoux

et al. 1974

The Journal of Cell Biology

215

112

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“…Subsequent ones will report on the effects of various treatments of the microsomes (5). Some of the findings made in these experiments have since been reported at a symposium (2), as part of other publications (1,7,18,39,40,44), and in abstract form (3).…”

Section: Introductionmentioning

confidence: 95%

Analytical Study of Microsomes and Isolated Subcellular Membranes From Rat Liver

Beaufay

Amar-Costesec

Thinessempoux

et al. 1974

The Journal of Cell Biology

237

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Rat liver microsomal fractions have been equilibrated in various types of linear density gradients. 15 fractions were collected and assayed for 27 constituents. As a result of this analysis microsomal constituents have been classified, in the order of increasing median density, into four groups labeled a, b, c, and d. Group a includes: monoamine oxidase, galactosyltransferase, 5'-nucleotidase, alkaline phosphodiesterase I, alkaline phosphatase, and cholesterol; group b: NADH cytochrome c reductase, NADPH cytochrome c reductase, aminopyrine demethylase, cytochrome b 5, and cytochrome P 450; group c: glucose 5-phosphatase, nucleoside diphosphatase, esterase, j3-glucuronidase, and glucuronyltransferase; group d: RNA, membrane-bound ribosomes, and some enzymes probably adsorbed on ribosomes: fumarase, aldolase, and glutamine synthetase. Analysis of the microsomal fraction by differential centrifugation in density gradient has further dissociated group a into constituents which sediment more slowly (monoamine oxidase and galactosyltransferase) than those of groups b and c, and 5'-nucleotidase, alkaline phosphodiesterase I, alkaline phosphatase, and the bulk of cholesterol which sediment more rapidly (group a2). The microsomal monoamine oxidase is attributed, at least partially, to detached fragments of external mitochondrial membrane. Galactosyltransferase belongs to the Golgi complex. Group a2 constituents are related to plasma membranes. Constituents of groups b and c and RNA belong to microsomal vesicles derived from the endoplasmic reticulum. These latter exhibit a noticeable biochemical heterogeneity and represent at the most 80% of microsomal protein, the rest being accounted for by particles bearing the constituents of groups a and some contaminating mitochondria, lysosomes, and peroxisomes. Attention is called to the operational meaning of microsomal subfractions and to their cytological complexity.

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“…This has been accomplished by differential centrifugation, a methodology developed in the 1940s and 1950s mainly by the cell biology group coordinated by Albert Claude, Christian de Duve and George Palade at the Rockefeller University. Their reviews should be consulted for descriptions of the early studies (De Duve 1971, Claude 1975, Palade 1975De Duve & Beaufay 1981. Their work established a baseline of important concepts: (a) polydispersity, which allowed the rules of sedimentation coefficients (Svedberg & Pedersen 1940), as defined for macromolecules, to be adapted for subcellular particle centrifugation; (b) biochemical homogeneity and unique localization, which resulted in the definition of marker enzymes; and (c) la-tency, which distinguished among enzymes confined in different membrane-bound compartments by the differences in time of release after membrane lyses (De Duve 1967).…”

Section: The Cell Fractionation Techniquementioning

confidence: 99%

Cell fractionation of parasitic protozoa: a review

Souza

Cunha-e-Silva

2003

Mem. Inst. Oswaldo Cruz

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Cell fractionation, a methodological strategy for obtaining purified organelle preparations, has been applied successfully to parasitic protozoa by a number of investigators. Here we present and discuss the work of several groups that have obtained highly purified subcellular fractions from trypanosomatids, Apicomplexa and trichomonads, and whose work have added substantially to our knowledge of the cell biology of these parasites.

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Tissue Fraction-Past and Present

Cited by 368 publications

References 97 publications

Analytical Study of Microsomes and Isolated Subcellular Membranes From Rat Liver

Analytical Study of Microsomes and Isolated Subcellular Membranes From Rat Liver

Analytical Study of Microsomes and Isolated Subcellular Membranes From Rat Liver

Cell fractionation of parasitic protozoa: a review

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