A New Method for Separation of Human Blood Components

Nakao, Makoto; Nakayama, Toshimasa; Kankura, Takako

doi:10.1038/newbio246094a0

Cited by 45 publications

(10 citation statements)

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“…Although Heller et al missed not only neutral proteases but also acid protease from erythrocyte membranes [8], we have established, by the present study, the existence of an erythrocyte membrane-bound acid protease, partly confirming the observation by Morrison and Neurath [l]. Leukocyte-free erythrocytes could also be isolated by passage through a sulphoethyl-cellulose column, as reported by Nakao et al [24]. When stroma suspensions were prepared from such erythrocytes and subjected to the studies on the protease, we obtained the results leading to the same conclusion as that described above.…”

Section: Sidedness Of Sealed Ghosts or Inside-out Vesicles And Specsupporting

confidence: 75%

An Acid Protease in Human Erythrocytes and Its Localization in the Inner Membrane

Murakami

Suzuki

Murachi

1979

European Journal of Biochemistry

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The isolation of erythrocytes of high purity from human blood was achieved by a combination of the two well established methods. Correlation between the proteolytic activity and the content of white blood cells in erythrocyte preparations of different purities was studied. The acid protease activity was recovered to a level comparable with the recovery of erythrocytes, while the neutral protease activity as detected by the release of acid-soluble peptides from hemoglobin or casein disappeared in proportion to the removal of white blood cells. An acid protease was solubilized from the membranes of the purified erythrocytes by the extraction with 1-butanol. The enzyme was active in a pH range from 2 to 4, and sensitive to pepstatin. It was named pH-3 protease after its pH optimum. Sealed ghosts with right-side-out membranes and inside-out vesicles with reverted membranes were prepared from the purified erythrocytes and compared with respect to pH-3 protease activity for its latency as well as its inactivation by tryptic digestion. The results obtained indicate that pH-3 protease is localized on the inner surface of erythrocyte membranes. The selfdigestion experiments at pH 4 using the sealed ghosts showed higher availability to pH-3 protease of spectrin and IVa protein than the other membrane proteins, also suggesting the localization of an acid protease in the inner membranes of erythrocytes.

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Section: Sidedness Of Sealed Ghosts or Inside-out Vesicles And Specsupporting

confidence: 75%

An Acid Protease in Human Erythrocytes and Its Localization in the Inner Membrane

Murakami

Suzuki

Murachi

1979

European Journal of Biochemistry

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“…Fresh heparinized blood samples (2-3 h after collection) were filtered through SE-celluloseSephadex G-25 mixture [5] in order to remove most leukocytes and platelets. The erythrocytes were washed in saline solution, hemolyzed by freezing and thawing and assayed for GSH-Px ac tivity at 37 °C according to the method of Beutler [3] using t-butyl hydroperoxide as substrate.…”

Section: Methodsmentioning

confidence: 99%

Red Cell Glutathione Peroxidase in Various Jewish Ethnic Groups in Israel

Golan¹,

Ezzer²,

Szienberg³

1980

Hum Hered

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Red cell glutathione peroxidase (GSH-Px) activity has been measured in samples from seven Jewish population groups in Israel. No significant differences were found between the various communities. The mean GSH-Px activity was similar to that observed by Beutler and Matsumoto in Jewish samples from the USA and Israel. According to a genetic model proposed by the above investigators, the sample was subdivided into three phenotypes with low, intermediate and high activity, suggesting gene frequencies of 0.5072 and 0.4928 for the GSH-Px^L and GSH-Px^H alleles, respectively.

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“…in PBS or with PBS alone. One day after the last injection, splenic RBCs were isolated from spleen cell suspensions on a Servacell/Sephadex column, as de scribed elsewhere [26], RBCs were adjusted to a concentration of 5X10*750)11 PBS and used as Ag. Spleen cell nuclei were prepared using nonionic detergents according to Zeller et al [27] with some modifications.…”

Section: Preparation O F Splenic Red Blood Cells (Rbcs) and Nuclei Fo Rmentioning

confidence: 99%

Murine Systemic Autoimmune Disease Induced by Mercuric Chloride: T Helper Cells Reacting to Self Proteins

Kubicka-Muranyi

Kremer²,

Rottmann

et al. 1996

Int Arch Allergy Immunol

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HgCl₂ induces a CD4+ T-cell-dependent systemic autoimmune disease in susceptible strains of rats and mice. In rats, autoreactive T cells were shown to be involved, whereas in mice, attention has focussed on the demonstration of ‘Hg-specific’ T cells. To clarify these seemingly different T cell involvements, T cells from B10.S mice treated with HgCl₂ for 1 or 8 weeks were analyzed for their capacity to mount anamnestic responses against various self antigens (Ags) which either contained Hg or did not. T cells from donors short-term treated with HgCl₂ failed to mount memory responses to Hg-free Ags, but mounted a significant response to HgCl₂ and also reacted with Hg-containing self Ags. Interestingly, T cells from donors long-term treated with HgCl₂ showed a different pattern of reactivity. They hardly reacted to HgCl₂ and reacted poorly to Hg-containing splenic proteins, but responded vigorously to nuclei and fibrillarin irrespective of whether these self constituents had been treated with HgCl₂ or not. Conceivably, the initial activation of T cells that recognize Hg in combination with nuclear self proteins, such as fibrillarin, eventually results in activation of T cells specific for the unaltered self proteins.

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A New Method for Separation of Human Blood Components

Cited by 45 publications

References 1 publication

An Acid Protease in Human Erythrocytes and Its Localization in the Inner Membrane

An Acid Protease in Human Erythrocytes and Its Localization in the Inner Membrane

Red Cell Glutathione Peroxidase in Various Jewish Ethnic Groups in Israel

Murine Systemic Autoimmune Disease Induced by Mercuric Chloride: T Helper Cells Reacting to Self Proteins

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