Characterization of the GTP-dependent activation of the superoxide-producing NADPH oxidase in a cell-free system of pig neutrophils

1993 ◽  
Vol 1178 (1) ◽  
pp. 73-80 ◽  
Author(s):  
Lixin Yu ◽  
Koichiro Takeshige ◽  
Hiroyuki Nunoi ◽  
Shigeki Minakami
Keyword(s):  
Nadph Oxidase ◽  
Free System ◽  
Cell Free System ◽  
A Cell

A somatic cell-derived system for studying both early and late mitotic events in vitro

1989 ◽  
Vol 94 (3) ◽  
pp. 449-462
Author(s):  
J. Nakagawa ◽  
G.T. Kitten ◽  
E.A. Nigg
Keyword(s):  
Chromatin Condensation ◽  
Nuclear Lamina ◽  
Free System ◽  
Cell Free System ◽  
Protein Substrates ◽  
Nuclear Envelope Breakdown ◽  
Biochemical Fractionation ◽  
A Cell

We describe a cell-free system for studying mitotic reorganization of nuclear structure. The system utilizes soluble extracts prepared from metaphase-arrested somatic chicken cells and supports both the disassembly and subsequent partial reassembly of exogenous nuclei. By fluorescence microscopy, biochemical fractionation, protein phosphorylation assays and electron microscopy, we show that chicken embryonic nuclei incubated in extracts prepared from metaphase-arrested chicken hepatoma cells undergo nuclear envelope breakdown, lamina depolymerization and chromatin condensation. These prophase-like events are strictly dependent on ATP and do not occur when nuclei are incubated in interphase extracts. Compared to interphase extracts, metaphase extracts show increased kinase activities toward a number of nuclear protein substrates, including lamins and histone H1; moreover, they specifically contain four soluble phosphoproteins of Mr 38,000, 75,000, 95,000 and 165,000. Following disassembly of exogenous nuclei in metaphase extracts, telophase-like reassembly of a nuclear lamina and re-formation of nuclear membranes around condensed chromatin can be induced by depletion of ATP from the extract. We anticipate that this reversible cell-free system will contribute to the identification and characterization of factors involved in regulatory and mechanistic aspects of mitosis.

Characterization of phospholipase D in a cell-free system of cultured cells derived from rat frontal cortex

1992 ◽  
Vol 595 (1) ◽  
pp. 12-16 ◽  
Author(s):  
Akira Nishida ◽  
Masami Shimizu ◽  
Yasunori Kanaho ◽  
Yoshinori Nozawa ◽  
Shigeto Yamawaki
Keyword(s):  
Frontal Cortex ◽  
Phospholipase D ◽  
Cultured Cells ◽  
Free System ◽  
Cell Free System ◽  
A Cell ◽  
Rat Frontal Cortex
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