Glucose-6-phosphate Dehydrogenase Isoenzymes in Blood Cells

Rozenszajn, L. A.; Kolman, Stanley D.; Shoham, D.; Gazith, J.

doi:10.1038/226862a0

Cited by 7 publications

(3 citation statements)

References 4 publications

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“…The definite differences in Rf among the three forms present at the extraporated zero concentration indicated that these forms had similar molecular sizes, but differed from each other in net charge [15]. This might explain the difficulty encountered in separating similar multiple forms of erythrocyte G6PD by gel filtration on Sephadex [4], Two or three iso zymes of G6PD in normal erythrocyte, reported by Rozenszajn et al [13], also showed no difference in the molecular size.…”

Section: Methodsmentioning

confidence: 72%

Three Interconvertible Forms of Glucose-6-Phosphate Dehydrogenase in Human Liver and Erythrocyte

Watanabe¹,

Taketa²,

Kosaka³

1972

Enzyme

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The separation of three electrophoretically distinct subcomponents of normal G6PD (type B^+) in human liver and erythrocyte has been achieved by means of disc electrophoresis. Molecular weights of these forms were found to be similar, and their resolution on disc electrophoresis appeared to be mainly due to the difference in net charge. The three forms were demonstrated to be interconvertible by treatment with sulfhydryl reagents

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Section: Methodsmentioning

confidence: 72%

Three Interconvertible Forms of Glucose-6-Phosphate Dehydrogenase in Human Liver and Erythrocyte

Watanabe¹,

Taketa²,

Kosaka³

1972

Enzyme

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“…Single colonies that grew inside the upper agar layer [type I colonies (11)] were isolated with a pasteur pipette at 6 or 7 days after seeding, after 1 ml of PBS had been added and the colonies that had grown on top of the agar (11) and were floating in the PBS had been removed. The isolated type I colonies were then smeared on a slide and the cells were stained for G6PD (17)(18)(19) by incubation for 2 hr at 370 in a solution that stained this enzyme. The cells were incubated under a coverslip with a solution containing 0.2 M D-glucose 6-phosphate, disodium salt (18).…”

Section: Methodsmentioning

confidence: 99%

“…The isolated type I colonies were then smeared on a slide and the cells were stained for G6PD (17)(18)(19) by incubation for 2 hr at 370 in a solution that stained this enzyme. The cells were incubated under a coverslip with a solution containing 0.2 M D-glucose 6-phosphate, disodium salt (18). Fifty cells were counted per colony.…”

Section: Methodsmentioning

confidence: 99%

Regulation of the induction of colonies in vitro by normal human lymphocytes.

Gerassi¹,

Sachs²

1976

Proc. Natl. Acad. Sci. U.S.A.

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Lymphocytes isolated from normal human peripheral blood can be induced to form colonies in vitro by incubation with the appropriate inducer. Phytohemagglutinin can induce colonies with T (thymus-derived) lymphocytes. Optimum colony formation with about two colonies per 102 peripheral blood lymphocytes was obtained by adding, in addition to phytohemagglutinin, autologous plasma, autologous red blood cells, and fresh L-glutamine or L-cystine. In the absence of these fresh amino acids, no colonies were formed at seeding levels below 105 cells per 35 mm petri dish. The addition of either of these amino acids gave a 10-fold decrease in the minimum number of cells that had to be seeded for colony formation. Lipopolysaccharide did not induce the formation of colonies, but enhanced the formation of T cell colonies by phytohemagglutinin. The mixing of lymphocytes from persons with and deficient in glucose-6-phosphate-dehydrogenase (EC 1. The development of a technique for the in vitro cloning in semi-solid medium of normal macrophages and granulocytes (1-4) has made it possible to elucidate the control of growth and differentiation of these two types of white blood cells and the changes in this control in myeloid leukemia and nonmalignant granulocyte diseases (5-8). The formation of macrophage and granulocyte colonies requires a specific protein inducer which we now call MGI (macrophage and granulocyte inducer) (6) but does not seem to require the addition of a foreign antigen. However, normal lymphocytes require antigenic stimulation for differentiation in mass culture in liquid medium (9, 10), and this suggested that normal lymphocytes could be cloned in semi-solid medium by adding the appropriate antigenic inducer.We have previously shown (11) that normal lymphocytes isolated from human peripheral blood and incubated with the appropriate inducer can form colonies in semi-solid medium.The inducers used were the T (thymus-derived) cell lectin phytohemagglutinin (PHA) and the T and B (bone-marrowderived) cell lectin pokeweed mitogen (PWM indicated that colony formation requires a critical concentration of some factors produced by the seeded cells in addition to the presence of the appropriate lectin (11). The present experiments were undertaken to clarify the specific factors required for optimum colony formation, to determine the clonal origin of the colonies, and to study the inducing effect of lipopolysaccharide (LPS) and its interaction with PHA and PWM. 2-Mercaptoethanol, which has been reported to induce the formation of B cell colonies in cultures of unfractionated cells from mouse hemapoietic organs (12), did not induce the formation of colonies with isolated human peripheral blood lymphocytes (11). MATERIALS AND METHODSCollection and Isolation of Peripheral Blood Lymphocytes. Peripheral blood from healthy adult volunteers was collected in citrate/phosphate/dextrose anticoagulant solution, and the plasma and buffy coat were separated. The cells were separated from the plasma by centrifugation and resuspe...

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Glucose‐6‐Phosphate Dehydrogenases

1979

Advances in Enzymology - And Related Areas of Molecular Biology

103

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Glucose-6-phosphate Dehydrogenase Isoenzymes in Blood Cells

Cited by 7 publications

References 4 publications

Three Interconvertible Forms of Glucose-6-Phosphate Dehydrogenase in Human Liver and Erythrocyte

Three Interconvertible Forms of Glucose-6-Phosphate Dehydrogenase in Human Liver and Erythrocyte

Regulation of the induction of colonies in vitro by normal human lymphocytes.

Glucose‐6‐Phosphate Dehydrogenases

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