Expression, purification, and characterization of a His6-tagged glycerokinase from Pichia farinosa for enzymatic cycling assays in mammalian cells

2010 ◽  
Vol 150 (3) ◽  
pp. 396-403 ◽  
Author(s):  
R. Janke ◽  
Y. Genzel ◽  
S. Freund ◽  
M.W. Wolff ◽  
H. Grammel ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Purification And Characterization ◽  
Enzymatic Cycling ◽  
Pichia Farinosa

scholarly journals Large scale purification and characterization of recombinant human autotaxin/lysophospholipase D from mammalian cells

BMB Reports ◽  
2010 ◽  
Vol 43 (8) ◽  
pp. 541-546 ◽  
Author(s):  
Yuanda Song ◽  
Emily Dilger ◽  
Jessica Bell ◽  
William A. Barton ◽  
Xianjun Fang
Keyword(s):  
Mammalian Cells ◽  
Large Scale ◽  
Purification And Characterization ◽  
Lysophospholipase D

scholarly journals Expression in mammalian cells, purification and characterization of recombinant human pancreatic ribonuclease

FEBS Letters ◽  
1993 ◽  
Vol 333 (3) ◽  
pp. 233-237 ◽  
Author(s):  
Nello Russo ◽  
Mena de Nigris ◽  
Antonietta Ciardiello ◽  
Alberto Di Donato ◽  
Giuseppe D'Alessio
Keyword(s):  
Mammalian Cells ◽  
Purification And Characterization ◽  
Pancreatic Ribonuclease ◽  
Expression In Mammalian Cells ◽  
Human Pancreatic Ribonuclease

scholarly journals Purification and Characterization of a Novel Physiological Substrate for Calcineurin in Mammalian Cells

1998 ◽  
Vol 273 (35) ◽  
pp. 22738-22744 ◽  
Author(s):  
Guy E. Groblewski ◽  
Mutsumi Yoshida ◽  
M. Julia Bragado ◽  
Stephan A. Ernst ◽  
Joseph Leykam ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Purification And Characterization

Purification and characterization of human phosphatidylserine synthases 1 and 2

2009 ◽  
Vol 418 (2) ◽  
pp. 421-429 ◽  
Author(s):  
Shiho Tomohiro ◽  
Ayako Kawaguti ◽  
Yukiyo Kawabe ◽  
Sakae Kitada ◽  
Osamu Kuge
Keyword(s):  
Feedback Control ◽  
Substrate Specificity ◽  
Mammalian Cells ◽  
Enzyme Assay ◽  
The Other ◽  
Significant Extent ◽  
Purification And Characterization ◽  
Intact Cells ◽  
Base Exchange

PS (phosphatidylserine) in mammalian cells is synthesized by two distinct base-exchange enzymes, PSS1 (PS synthase 1) and PSS2, which are responsible for the conversion of PC (phosphatidylcholine) and PE (phosphatidylethanolamine) respectively into PS in intact cells. The PS synthesis in cultured mammalian cells is inhibited by exogenous PS, and this feedback control occurs through inhibition of PSSs by PS. In the present study, we purified epitope-tagged forms of human PSS1 and PSS2. The purified PSS2 was shown to catalyse the conversion of PE, but not PC, into PS, this being consistent with the substrate specificity observed in intact cells. On the other hand, the purified PSS1 was shown to catalyse the conversion of both PC and PE into PS, although PSS1 in intact cells had been shown not to contribute to the conversion of PE into PS to a significant extent. Furthermore, we found that the purified PSS2, but not the purified PSS1, was inhibited on the addition of PS to the enzyme assay mixture, raising the possibility that there was some difference between the mechanisms of the inhibitory actions of PS towards PSS1 and PSS2.

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