Baculovirus expression, purification, and characterization of human protein phosphatase 2A catalytic subunits α and β

Ikehara, Tsuyoshi; Shinjo, Fukiko; Ikehara, Satsuki; Imamura, Shihoko; Yasumoto, Takeshi

doi:10.1016/j.pep.2005.06.002

Cited by 37 publications

(44 citation statements)

References 34 publications

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“…The activity against T220 was lower than 10 μmol/min/mg and the specific activities against the other phosphopeptides were almost zero. According to a previous study 20 for PP2A activity against the RRAT(p)VA peptide, PP2A Cα had a specific activity of 10.89 ± 2.44 μmol/min/mg, which was reconfirmed in this study when PP2A Cα was included as a positive control. Therefore, the specific activities of PP2A for S460 and S454 are comparable to that of the RRAT(p)VA peptide.…”

Section: Notessupporting

confidence: 80%

“…The other 5 phosphopeptides (T220, S240, S245, S250, and S260) are located in the linker region of SMAD2. We evaluated the specific activity of SCP1 and PP2A Cα with the 13 phosphopeptides of SMAD2 under reaction conditions originating from previous reports 2,20 to compare the relative activities. SCP1 had the highest specific activity against S250 and the second highest specific activity against S458 ( Fig.…”

Section: Notesmentioning

confidence: 99%

“…PP2A Cα-catalyzed dephosphorylation of the phosphorylated substrate was performed as previously described. 20 Assays were conducted at 37°C in a buffer consisting of 40 mM Tris-HCl (pH 8.4), 34 mM MgCl 2 , 4 mM EDTA, 2 mM DTT, 0.05 mg/mL BSA, 5 μM-1 mM phosphopeptides, and 5-60 ng of PP2A Cα. Phosphate release was quantitated by using a malachite green-based colorimetric assay for inorganic phosphate by measuring the absorbance at 620 nm.…”

Section: -261mentioning

confidence: 99%

See 2 more Smart Citations

Selective Dephosphorylation by SCP1 and PP2A in Phosphorylated Residues of SMAD2

Hong¹,

Sung²,

Lee³

et al. 2014

Bulletin of the Korean Chemical Society

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Protein phosphatase is important for cellular events, and a family of protein phosphatases, the so-called C-terminal domain (CTD) of RNA polymerase II (RNAPII) phosphatases, has recently attracted attention. The CTD is the largest subunit of RNAPII and consists of a tandem repeated heptapeptide (Y 1 S 2 P 3 T 4 S 5 P 6 S 7 ).1 Seven active CTD phosphatases in the human genome are known and share the same catalytic domain architecture and DXDX(T/V) active site motif.2 Small CTD phosphatase 1 (SCP1) dephosphorylates the fifth phosphorylated serine of Y 1 S 2 P 3 T 4 S 5 P 6 S 7 in the CTD 2 and has been renamed CTD small phosphatase 1 (CTDSP1). Small CTD phosphatase 2 (SCP2) and small CTD phosphatase 3 (SCP3) have a similar sequence, threedimensional structure, and biochemical function to SCP1.

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Section: Notessupporting

confidence: 80%

Section: Notesmentioning

confidence: 99%

Section: -261mentioning

confidence: 99%

See 1 more Smart Citation

Selective Dephosphorylation by SCP1 and PP2A in Phosphorylated Residues of SMAD2

Hong¹,

Sung²,

Lee³

et al. 2014

Bulletin of the Korean Chemical Society

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“…Reagents-His 8 -tagged Xenopus PP2A C␤ was produced in High Five cells as described previously (28) and purified by modified ethanol precipitation (29) and affinity chromatography with nickel-nitrilotriacetic acid resin. (Qiagen).…”

Section: Methodsmentioning

confidence: 99%

Control of DNA Replication by the Nucleus/Cytoplasm Ratio in Xenopus

Murphy

Michael

2013

Journal of Biological Chemistry

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Background:The duration of S phase in Xenopus is controlled by the nucleus/cytoplasm ratio through unknown mechanisms. Results: Modulation of protein phosphatase 2A (PP2A) levels controls origin firing at high nucleus/cytoplasm. Conclusion: Depletion of PP2A and its B55␣ subunit at high nucleus/cytoplasm causes an extension of S phase. Significance: This mechanism may influence cell cycle remodeling that occurs in early embryogenesis.

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“…PP2Ac was kindly provided by Dr. Tsuyoshi Ikehara (38) and cloned into the pRK5-FLAG vector. PKC␤II-T634A/T638A/T641A, PKC␤II-S660A, and PKC␤II (S660E) were kindly provided by Dr. Alexandra Newton (36).…”

Section: Methodsmentioning

confidence: 99%

Regulation of Protein Kinase C Inactivation by Fas-associated Protein with Death Domain

Cheng

Wang

Zhang

et al. 2012

Journal of Biological Chemistry

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Background: PKC is extremely important for a wide array of cellular processes. However, its inactivation is poorly understood. Results: FADD deficiency or phosphoryl-mimicking mutation (FADD-D) leads to accumulation of phosphorylated PKC and sustained signaling. Conclusion:The apoptotic adapter FADD is required for PKC dephosphorylation, degradation and signaling inactivation and may be regulated by its phosphorylation. Significance: FADD is critical for PKC dephosphorylation, stability, and signaling termination.

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Baculovirus expression, purification, and characterization of human protein phosphatase 2A catalytic subunits α and β

Cited by 37 publications

References 34 publications

Selective Dephosphorylation by SCP1 and PP2A in Phosphorylated Residues of SMAD2

Selective Dephosphorylation by SCP1 and PP2A in Phosphorylated Residues of SMAD2

Control of DNA Replication by the Nucleus/Cytoplasm Ratio in Xenopus

Regulation of Protein Kinase C Inactivation by Fas-associated Protein with Death Domain

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