A Newly Synthesized Selective Casein Kinase I Inhibitor, N-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide, and Affinity Purification of Casein Kinase I from Bovine Testis

Chijiwa, Takashi; Hagiwara, Masatoshi; Hidaka, Hiroyoshi

doi:10.1016/s0021-9258(18)83679-1

Cited by 153 publications

(13 citation statements)

References 22 publications

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“…Fig. 8 demonstrates that this protein kinase is inhibited by both CKI-7 and CKI-8, specific inhibitors of CKI activity (28). The IC50 value for CKI-7 inhibition of the synaptic vesicle CKI is 12 ~M, somewhat lower than the value reported for a-CKI (78).…”

Section: Isolation and Characterization Of The Synaptic Vesicle Associated 36-kd Ckimentioning

confidence: 81%

A phosphatidylinositol 4,5-bisphosphate-sensitive casein kinase I alpha associates with synaptic vesicles and phosphorylates a subset of vesicle proteins.

Größ

Hoffman

Fisette

et al. 1995

The Journal of Cell Biology

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Abstract. In interphase cells, a-casein kinase I (et-CKI) is found associated with cytosolic vesicular structures, the centrosome, and within the nucleus. To identify the specific vesicular structures with which a-CKI is associated, established cell lines and primary rat neurons were immunofluorescently labeled with an antibody raised to a-CKI. In nonneuronal cells, a-CKI colocalizes with vesicular structures which align with microtubules and are partially coincident with both Golgi and endoplasmic reticulum markers. In neurons, o~-CKI colocalizes with synaptic vesicle markers. When synaptic vesicles were purified from rat brain, they were highly enriched in a CKI, based on activity and immunoreactivity. The synaptic vesicle-associated CKI is an extrinsic kinase and was eluted from synaptic vesicles and purified. This purified CKI has properties most similar to o~-CKI. When the activities of casein kinase I or II were specifically inhibited on isolated synaptic vesicles, CKI was shown to phosphorylate a specific subset of vesicle proteins, one of which was identified as the synaptic vesicle-specific protein SV2. As with ot-CKI, the synaptic vesicle CKI is inhibited by phosphatidylinositol 4,5-bisphosphate (PIP2). However, synthesis of PIP2 was detected only in plasma membranecontaining fractions. Therefore, PIP2 may spatially regulate CKI. Since PIP2 synthesis is required for secretion, this inhibition of CKI may be important for the regulation of secretion. CELLS transport proteins and lipids throughout their cytoplasm using small vesicles containing specific integral membrane proteins (2-5, 13, 24, 35, 38, 52, 58, 62, 72-74). This transport system is microtubule dependent and requires motor proteins (16,19,31). In this process, vesicles bud from donor membranes and are shunted to specific acceptor membranes where the vesicles fuse and release their contents. These general events can be divided into constituitive and regulated pathways (13,17,24,35,41,65,(72)(73)(74). Although distinct, the two pathways appear to have similar components in common. A typical constitutive pathway would be the shuttling of vesicles within and between the endoplasmic reticulum, the Golgi organelle, and the plasma membrane. The movement and CaE+-dependent fusion of synaptic vesicles provides an example of a regulated secretory pathway.Stidhof and Jahn (74) physin and synaptobrevin, and two proteins integral to the plasma membrane, syntaxin and neurexin are involved in docking (4,5,10,11,26,62,68,75). Synaptotagmin, another vesicle integral membrane protein, is thought to act in both the positioning of vesicles at the plasma membrane and the Ca2+-dependent step of the fusion process (12,22,34,37,55,60,61). Finally, the SNAPs (N-ethylmaleimidesensitive factor-associated proteins) have been linked to vesicle targeting and fusion (11,15,35,71,77).Within each of these stages, the myriad of processes involved are extensively regulated. Evidence exists showing that G-proteins (16-18, 23, 41, 42), phosphoinositide kinases (23, 45-48, 69),...

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Section: Isolation and Characterization Of The Synaptic Vesicle Associated 36-kd Ckimentioning

confidence: 81%

A phosphatidylinositol 4,5-bisphosphate-sensitive casein kinase I alpha associates with synaptic vesicles and phosphorylates a subset of vesicle proteins.

Größ

Hoffman

Fisette

et al. 1995

The Journal of Cell Biology

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“…6 E). Furthermore, the enhanced Tcf3 phosphorylation by CK1⑀ is partially inhibited when CKI-7 (IC 50 , 10-30 M; purchased from Seikagaku Corporation), a selective inhibitor of CK1 activity (Chijiwa et al, 1989), is included in the kinase reaction. The association between Tcf3 and GKS3/CK1⑀ represents bona fide in vivo interactions, since both endogenous Xenopus GSK3 and CK1e immunoprecipitated with an antimyc antibody from extracts made from embryos injected with myc 6 -xTcf3 mRNA (Fig.…”

Section: Tcf3 Phosphorylation By Gsk3 and Ck1⑀mentioning

confidence: 99%

Physiological regulation of β-catenin stability by Tcf3 and CK1ϵ

Lee

Salic

Kirschner

2001

The Journal of Cell Biology

142

121

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The wnt pathway regulates the steady state level of β-catenin, a transcriptional coactivator for the Tcf3/Lef1 family of DNA binding proteins. We demonstrate that Tcf3 can inhibit β-catenin turnover via its competition with axin and adenomatous polyposis for β-catenin binding. A mutant of β-catenin that cannot bind Tcf3 is degraded faster than the wild-type protein in Xenopus embryos and extracts. A fragment of β-catenin and a peptide encoding the NH2 terminus of Tcf4 that block the interaction between β-catenin and Tcf3 stimulate β-catenin degradation, indicating this interaction normally plays an important role in regulating β-catenin turnover. Tcf3 is a substrate for both glycogen synthase kinase (GSK) 3 and casein kinase (CK) 1ε, and phosphorylation of Tcf3 by CKIε stimulates its binding to β-catenin, an effect reversed by GSK3. Tcf3 synergizes with CK1ε to inhibit β-catenin degradation, whereas CKI-7, an inhibitor of CK1ε, reduces the inhibitory effect of Tcf3. Finally, we provide evidence that CK1ε stimulates the binding of dishevelled (dsh) to GSk3 binding protein (GBP) in extracts. Along with evidence that a significant amount of Tcf protein is nonnuclear, these findings suggest that CK1ε can modulate wnt signaling in vivo by regulating both the β-catenin-Tcf3 and the GBP-dsh interfaces.

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“…The Km value for ATP is similar to those reported for other casein kinase I preparations (usually 7-25 uM; Tuazon and Traugh, 1991). So is the IC50 value for CKI-7 (9.5 ,uM for casein kinase I from bovine testis; Chijiwa et al, 1989). However, the plant enzyme is not stimulated by univalent cations (Table 2).…”

Section: Discussionmentioning

confidence: 99%

“…Because of the still lower abundance of the plant enzyme, this problem was even more pronounced, and significant losses occurred during classical column procedures to a degree that abolished any resulting purification. Therefore we adapted the affinity procedure on an immobilized casein kinase I-specific inhibitor, CKI-7, developed by Chijiwa et al (1989).…”

Section: Calculation Of Kinetic Parametersmentioning

confidence: 99%

Purification and characterization of casein kinase I from broccoli

Klimczak

Cashmore

1993

Biochemical Journal

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Casein kinase I from broccoli was purified approximately 65,000-fold by chromatography on phosphocellulose, phenyl-Sepharose, CM-Sephacel, and affinity chromatography on N-(2-aminoethyl)-5-chloroisoquinolone-8-sulphonamide (CKI-7)-Sepharose. The catalytic subunit of casein kinase I was identified as a 36-38 kDa polypeptide doublet by using the technique of activity gel assay after SDS/PAGE with casein as a gel-incorporated substrate. A silver-stained polypeptide doublet of the same molecular mass constituted at least 95% of the protein in the final preparation, corresponding to a specific activity of approximately 1800 nmol/min per mg of protein. The enzyme was found to be a monomer by gel filtration and glycerol gradient sedimentation; the native molecular mass was calculated to be 34.2 kDa. These characteristics, as well as other essential features of plant casein kinase I activity, such as substrate specificity and sensitivity to inhibitors, were found to be similar to those established for animal casein kinase I. Broccoli casein kinase I showed weak immunological cross-reactivity with antibodies raised against bovine casein kinase I.

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A Newly Synthesized Selective Casein Kinase I Inhibitor, N-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide, and Affinity Purification of Casein Kinase I from Bovine Testis

Cited by 153 publications

References 22 publications

A phosphatidylinositol 4,5-bisphosphate-sensitive casein kinase I alpha associates with synaptic vesicles and phosphorylates a subset of vesicle proteins.

A phosphatidylinositol 4,5-bisphosphate-sensitive casein kinase I alpha associates with synaptic vesicles and phosphorylates a subset of vesicle proteins.

Physiological regulation of β-catenin stability by Tcf3 and CK1ϵ

Purification and characterization of casein kinase I from broccoli

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