Purification of an NADPH‐dependent diaphorase from membrane of DMSO‐induced differentiated human promyelocytic leukemia HL‐60 cells

Otsuka-Murakami, Hidetsugu; Nisimoto, Yukio

doi:10.1016/0014-5793(95)00183-a

Cited by 3 publications

(3 citation statements)

References 15 publications

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“…7 The gels were incubated at 37°C for 30 min in the reaction mixture (0.4 mM NADPH, 8 mM MgCl 2 , 0.5 mM piodonitrotetrazolium violet and 10% glycerol in 50 mM phosphate buffer (pH 7.0)). 8 The incubation was stopped by washing in ice-cold buffer (50 mM phosphate buffer, pH 7.0).…”

Section: Nadph-diaphorase Activity Staining and Western Blot Analysismentioning

confidence: 99%

Nitric oxide participates in cataract development in selenite-treated rats

Ito¹,

Nabekura²,

Takeda³

et al. 2001

Current Eye Research

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Section: Nadph-diaphorase Activity Staining and Western Blot Analysismentioning

confidence: 99%

Nitric oxide participates in cataract development in selenite-treated rats

Ito¹,

Nabekura²,

Takeda³

et al. 2001

Current Eye Research

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“…Isolated rat hepatocytes in culture show all signs of rapid dedifferentiation together with the translocation of ZnBP into the nucleus. But this relationship is more than circumstantial as indicated by the fact that DMSO not only inhibits dedifferentiation of hepatocytes (Ai et al, 1995;Kuliczkowski et al, 1995;Otsuka-Murakami and Nishimoto, 1995), but also abolishes completely over a long time the translocation of ZnBP from the cytoplasm to the nucleus. It should also be noted, that the amount of ZnBP in human breast cancer is dramatically increased (Tsitsiloni et al, 1998), an observation we could confirm for hepatocarcinoma cells (unpublished results).…”

Section: Discussionmentioning

confidence: 99%

Factors involved in the cell density-dependent regulation of nuclear/cytoplasmic distribution of the 11.5-kDa Zn2+-binding protein (parathymosin-α) in rat hepatocytes

Trompeter¹,

Schiermeyer²,

Blankenburg³

et al. 1999

Journal of Cell Science

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Although the 11.5 kDa Zn(2+)-binding protein (ZnBP, parathymosin-alpha) possesses a functional bipartite nuclear localization signal it was found in most tissues in the cytoplasm. The cultivation of freshly isolated rat hepatocytes for 24 hours under standard conditions was associated with an almost complete translocation of ZnBP from the cytoplasm to the nuclei. Here we demonstrate, that this translocation is negatively correlated with cell density. The translocation of ZnBP to the nucleus can be inhibited or abolished by inhibitors of protein synthesis (cycloheximide) or transcription (actinomycin D). Moreover, cycloheximide can induce a relocation of ZnBP to the cytoplasm when applied after the appearance of ZnBP in the nuclei. DMSO, an inhibitor of dedifferentiation of cultured hepatocytes, abolishes also the translocation of ZnBP into the nucleus. Thinly seeded cells keep their ZnBP in the cytoplasm if they are co-cultured with plasma membranes from Morris MH7777 hepatoma cells or antibodies against E-cadherin indicating the involvement of cell adhesion proteins. We have enriched a protein from the cytosol of fresh hepatocytes which inhibits the translocation of ZnBP, but not that of albumin-NLS into the nucleus in a permeabilized cell system. Such an activity could not be found in the cytoplasm of permanent cell lines which harbor ZnBP only in the nucleus. A model for the regulation of the nuclear import of ZnBP is proposed.

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“…Several groups have reported that an additional flavin containing dehydrogenase from rabbit peritoneal neutrophils and pig neutrophils is able to enhance superoxide production in a cell‐free system [10–12]. Nisimoto et al [13, 14]have purified an NADPH‐cytochrome c reductase from human neutrophils and HL‐60 cells but did not observe a marked stimulation of superoxide generation by the purified reductase.…”

Section: Introductionmentioning

confidence: 99%

Papain proteolysis releases a soluble NADPH dependent diaphorase activity from bovine neutrophil membranes

1998

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An NADPH dependent cytochrome c reductase has been purified from resting bovine neutrophil membranes. A high degree of purification, approaching homogeneity, is indicated by the presence of a single 75 kDa protein band on silver stained SDS-PAGE (10%). The purified protein catalyzes as well an NADPH dependent reduction of iodonitrotetrazolium violet (INT). Limited papain digestion of the purified preparation produces a 65 kDa product which retains both enzymatic activities. In a similar fashion papain digestion of the plasma membrane bound protein generates a fully active soluble NADPH dependent INT and cytochrome c reductase preparation (65 kDa). Proteolytic cleavage would appear to occur at a protein-membrane anchor remote from the proteins catalytic site. The cytochrome c reductase acts independently of the O 3 2 generating cytochrome b SSV , a leukocyte plasma membrane protein which also catalyzes an NADPH dependent INT reduction.z 1998 Federation of European Biochemical Societies.

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Purification of an NADPH‐dependent diaphorase from membrane of DMSO‐induced differentiated human promyelocytic leukemia HL‐60 cells

Cited by 3 publications

References 15 publications

Nitric oxide participates in cataract development in selenite-treated rats

Nitric oxide participates in cataract development in selenite-treated rats

Factors involved in the cell density-dependent regulation of nuclear/cytoplasmic distribution of the 11.5-kDa Zn2+-binding protein (parathymosin-α) in rat hepatocytes

Papain proteolysis releases a soluble NADPH dependent diaphorase activity from bovine neutrophil membranes

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