Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells.

Cohen, Carl M.; Kalish, Douglas I.; Jacobson, Bernard; Branton, Daniel

doi:10.1083/jcb.75.1.119

Cited by 123 publications

(63 citation statements)

References 38 publications

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“…The sucrose density gradient membranes contained twice as much contaminating mitochondria enzyme and three times as much granule enzyme . It should be noted that these data provide only a rough estimation of purity in view of the fact that enzymatic functions in membranes attached on polylysine beads may be different from those in free membranes (5).…”

Section: Purity Of the Membrane Preparation Iodination Studiesmentioning

confidence: 99%

“…(5). In a plastic tube with a cap, 0.2-0.3 ml of the beads was incubated in 0.7-0 .8 ml of an individual reaction medium while constantly inverting the tube with a rotating apparatus in a dry incubator at 37°C .…”

Section: Enzymatic Assaysmentioning

confidence: 99%

“…The assay methods for Na,KATPase and cytochrome oxidase were those described by Cohen et al . (5). ,ß-Glucuronidase was measured according to the method of Stahl and Touster (30) .…”

Section: Enzymatic Assaysmentioning

confidence: 99%

“…(5). Polylysine beads were prepared with polyacrylamide beads as described by Jacobson and Branton (10) .…”

Section: Isolation Of Platelet Membranes With Polylysine Beadsmentioning

confidence: 99%

“…The platelet plasma membrane also appears to possess specific surface receptors for certain physiologic stimulatory agents such as ADP (17), epinephrine (20), thrombin (33), and prostaglandins (16) fractions of isolated membranes from subcellular components such as mitochondria and granules . Recently, Jacobson and Branton (10) developed a novel method using polylysine-coated beads for the isolation of erythrocyte membranes and HeLa cell membranes (5) . With this method, surface membranes of cells attach to the polylysine beads and, after cell disruption, relatively pure preparations of plasma membranes remain attached to the beads .…”

mentioning

confidence: 99%

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Isolation of human platelet plasma membranes with polylysine beads.

Kinoshita¹,

Nachman²,

Minick³

1979

The Journal of Cell Biology

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Human platelet plasma membranes were isolated with polylysine beads according to the technique developed by Jacobson and Branton (1977, Science [Wash. D. C.] 195:302--304). Lactoperoxidase-catalyzed surface iodination revealed that ninefold greater 125I specific activity was associated with the membranes isolated on beads than with whole platelets. Enrichment in the bead membrane preparation of the activities of membrane marker enzymes, bis(p-nitrophenyl)phosphate phosphodiesterase and Na,K-ATPase, was 8.0 and 4.4, respectively. Contamination with enzymes of other organelles, cytochrome oxidase and beta-glucuronidase, was relatively low as compared with membranes isolated by sucrose gradient centrifugation. Analysis by SDS polyacrylamide gel electrophoresis showed that a full complement of surface glycoproteins was present on the membranes isolated with polylysine beads. The polylysine bead technique is a rapid, reproducible and efficient method for the preparation of relatively pure platelet plasma membranes.

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Section: Purity Of the Membrane Preparation Iodination Studiesmentioning

confidence: 99%

Section: Enzymatic Assaysmentioning

confidence: 99%

“…The assay methods for Na,KATPase and cytochrome oxidase were those described by Cohen et al . (5). ,ß-Glucuronidase was measured according to the method of Stahl and Touster (30) .…”

Section: Enzymatic Assaysmentioning

confidence: 99%

“…(5). Polylysine beads were prepared with polyacrylamide beads as described by Jacobson and Branton (10) .…”

Section: Isolation Of Platelet Membranes With Polylysine Beadsmentioning

confidence: 99%

mentioning

confidence: 99%

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Isolation of human platelet plasma membranes with polylysine beads.

Kinoshita¹,

Nachman²,

Minick³

1979

The Journal of Cell Biology

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Functional Coating of Porous Silica Microparticles with Native Biomembranes towards Portable Flow‐Through Biochemical Microreactors

Tutuş

Kaufmann

Weiss

et al. 2012

Adv Funct Materials

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In biological cells, various transmembrane enzymes function as highly effective chemical reactors confined in space with characteristic length scales of tens of nanometers to micrometer. However, it is still challenging to quantitatively confine membranes in compact reactor platforms without losing their biochemical functions. Here, a simple and straightforward strategy towards the fabrication of a new flow‐through reactor by the functional coating of porous silica microparticles with sarcoplasmic reticulum membranes is described. After a short incubation, the membranes achieve the homogeneous, full coverage of the particle surface, spanning across pores with the diameter of about 100 nm. By using the underlying pores as cavity reservoirs, transmembrane enzyme (Ca2+‐ATPase) in the membrane retains their capability of ATP hydrolysis. This enables us to confine 1.1 m2 of native membranes containing a large amount of Ca2+‐ATPase (approx. 10 nmol) in a column‐packaged, flow‐through reactor with merely 1.8 mL volume, which cannot be achieved by the reconstitution of proteins in artificial lipid membranes or condensation of membranes in suspensions. The distinct functional levels corresponding to different reaction buffers can be reproduced even after many buffer exchanges over 14 days, confirming the stability and reproducibility of the membrane‐particle hybrid reactors.

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Mechanistic Studies on Peroxide Activation by a Water‐Soluble Iron(III)–Porphyrin: Implications for OO Bond Activation in Aqueous and Nonaqueous Solvents

Wolak

Eldik

2007

Chemistry A European J

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The reactions of a water-soluble iron(III)-porphyrin, [meso-tetrakis(sulfonatomesityl)porphyrinato]iron(III), [Fe(III)(tmps)] (1), with m-chloroperoxybenzoic acid (mCPBA), iodosylbenzene (PhIO), and H(2)O(2) at different pH values in aqueous methanol solutions at -35 degrees C have been studied by using stopped-flow UV/Vis spectroscopy. The nature of the porphyrin product resulting from the reactions with all three oxidants changed from the oxo-iron(IV)-porphyrin pi-cation radical [Fe(IV)(tmps(*+))(O)] (1(++)) at pH<5.5 to the oxo-iron(IV)-porphyrin [Fe(IV)(tmps)(O)] (1(+)) at pH>7.5, whereas a mixture of both species was formed in the intermediate pH range of 5.5-7.5. The observed reactivity pattern correlates with the E degrees' versus pH profile reported for 1, which reflects pH-dependent changes in the relative positions of E degrees'(Fe(IV)/Fe(III) ) and E degrees'(P(*+)/P) for metal- and porphyrin-centered oxidation, respectively. On this basis, the pH-dependent redox equilibria involving 1(++) and 1(+) are suggested to determine the nature of the final products that result from the oxidation of 1 at a given pH. The conclusions reached are extended to water-insoluble iron(III)-porphyrins on the basis of literature data concerning the electrochemical and catalytic properties of [Fe(III)(P)(X)] species in nonaqueous solvents. Implications for mechanistic studies on [Fe(P)]-catalyzed oxidation reactions are briefly addressed.

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Membrane isolation on polylysine-coated beads. Plasma membrane from HeLa cells.

Cited by 123 publications

References 38 publications

Isolation of human platelet plasma membranes with polylysine beads.

Isolation of human platelet plasma membranes with polylysine beads.

Functional Coating of Porous Silica Microparticles with Native Biomembranes towards Portable Flow‐Through Biochemical Microreactors

Mechanistic Studies on Peroxide Activation by a Water‐Soluble Iron(III)–Porphyrin: Implications for OO Bond Activation in Aqueous and Nonaqueous Solvents

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