Identification and characterization of novel components of a Ca 2+/calmodulin-dependent protein kinase cascade in HeLa cells

Yumi Ishikawa; Hiroshi Tokumitsu; Hiroyuki Inuzuka; Maki Murata-Hori; Hiroshi Hosoya; Ryoji Kobayashi

doi:10.1016/S0014-5793(03)00817-2

Identification and characterization of novel components of a Ca ²⁺/calmodulin-dependent protein kinase cascade in HeLa cells

Yumi Ishikawa, Hiroshi Tokumitsu, Hiroyuki Inuzuka, Maki Murata-Hori, Hiroshi Hosoya, Ryoji Kobayashi

Research output: Contribution to journal › Article

27 Citations (Scopus)

Abstract

In this report, we cloned a novel calmodulin-kinase (CaM-KIδ) from HeLa cells and characterized its activation mechanism. CaM-KIδ exhibits Ca²⁺/CaM-dependent activity that is enhanced (∼30-fold) in vitro by phosphorylation of its Thr180 by CaM-K kinase (CaM-KK)α, consistent with detection of CaM-KIδ-activating activity in HeLa cells. We also identified a novel CaM-KKβ isoform (CaM-KKβ-3) in HeLa cells whose activity was highly Ca²⁺/CaM-independent. Transiently expressed CaM-KIδ exhibited enhanced protein kinase activity in HeLa cells without ionomycin stimulation. This sustained activation of CaM-KIδ was completely abolished by Thr180Ala mutation and inhibited by CaM-KK inhibitor, STO-609, indicating a functional CaM-KK/CaM-KIδ cascade in HeLa cells.

Original language	English
Pages (from-to)	57-63
Number of pages	7
Journal	FEBS Letters
Volume	550
Issue number	1-3
DOIs	https://doi.org/10.1016/S0014-5793(03)00817-2
Publication status	Published - Aug 28 2003

Fingerprint

Keywords

Ca
CaM-K kinase
CaM-kinase cascade
CaM-kinaseδ

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Cite this

Ishikawa, Y., Tokumitsu, H., Inuzuka, H., Murata-Hori, M., Hosoya, H., & Kobayashi, R. (2003). Identification and characterization of novel components of a Ca ²⁺/calmodulin-dependent protein kinase cascade in HeLa cells. FEBS Letters, 550(1-3), 57-63. https://doi.org/10.1016/S0014-5793(03)00817-2

@article{eaad7a36b7fe44958ef3b0ded63f00a8,

title = "Identification and characterization of novel components of a Ca 2+/calmodulin-dependent protein kinase cascade in HeLa cells",

abstract = "In this report, we cloned a novel calmodulin-kinase (CaM-KIδ) from HeLa cells and characterized its activation mechanism. CaM-KIδ exhibits Ca2+/CaM-dependent activity that is enhanced (∼30-fold) in vitro by phosphorylation of its Thr180 by CaM-K kinase (CaM-KK)α, consistent with detection of CaM-KIδ-activating activity in HeLa cells. We also identified a novel CaM-KKβ isoform (CaM-KKβ-3) in HeLa cells whose activity was highly Ca2+/CaM-independent. Transiently expressed CaM-KIδ exhibited enhanced protein kinase activity in HeLa cells without ionomycin stimulation. This sustained activation of CaM-KIδ was completely abolished by Thr180Ala mutation and inhibited by CaM-KK inhibitor, STO-609, indicating a functional CaM-KK/CaM-KIδ cascade in HeLa cells.",

keywords = "Ca, CaM-K kinase, CaM-kinase cascade, CaM-kinaseδ",

author = "Yumi Ishikawa and Hiroshi Tokumitsu and Hiroyuki Inuzuka and Maki Murata-Hori and Hiroshi Hosoya and Ryoji Kobayashi",

year = "2003",

month = aug

day = "28",

doi = "10.1016/S0014-5793(03)00817-2",

language = "English",

volume = "550",

pages = "57--63",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "Elsevier",

number = "1-3",

}

TY - JOUR

T1 - Identification and characterization of novel components of a Ca 2+/calmodulin-dependent protein kinase cascade in HeLa cells

AU - Ishikawa, Yumi

AU - Tokumitsu, Hiroshi

AU - Inuzuka, Hiroyuki

AU - Murata-Hori, Maki

AU - Hosoya, Hiroshi

AU - Kobayashi, Ryoji

PY - 2003/8/28

Y1 - 2003/8/28

N2 - In this report, we cloned a novel calmodulin-kinase (CaM-KIδ) from HeLa cells and characterized its activation mechanism. CaM-KIδ exhibits Ca2+/CaM-dependent activity that is enhanced (∼30-fold) in vitro by phosphorylation of its Thr180 by CaM-K kinase (CaM-KK)α, consistent with detection of CaM-KIδ-activating activity in HeLa cells. We also identified a novel CaM-KKβ isoform (CaM-KKβ-3) in HeLa cells whose activity was highly Ca2+/CaM-independent. Transiently expressed CaM-KIδ exhibited enhanced protein kinase activity in HeLa cells without ionomycin stimulation. This sustained activation of CaM-KIδ was completely abolished by Thr180Ala mutation and inhibited by CaM-KK inhibitor, STO-609, indicating a functional CaM-KK/CaM-KIδ cascade in HeLa cells.

AB - In this report, we cloned a novel calmodulin-kinase (CaM-KIδ) from HeLa cells and characterized its activation mechanism. CaM-KIδ exhibits Ca2+/CaM-dependent activity that is enhanced (∼30-fold) in vitro by phosphorylation of its Thr180 by CaM-K kinase (CaM-KK)α, consistent with detection of CaM-KIδ-activating activity in HeLa cells. We also identified a novel CaM-KKβ isoform (CaM-KKβ-3) in HeLa cells whose activity was highly Ca2+/CaM-independent. Transiently expressed CaM-KIδ exhibited enhanced protein kinase activity in HeLa cells without ionomycin stimulation. This sustained activation of CaM-KIδ was completely abolished by Thr180Ala mutation and inhibited by CaM-KK inhibitor, STO-609, indicating a functional CaM-KK/CaM-KIδ cascade in HeLa cells.

KW - Ca

KW - CaM-K kinase

KW - CaM-kinase cascade

KW - CaM-kinaseδ

UR - http://www.scopus.com/inward/record.url?scp=0142039017&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0142039017&partnerID=8YFLogxK

U2 - 10.1016/S0014-5793(03)00817-2

DO - 10.1016/S0014-5793(03)00817-2

M3 - Article

C2 - 12935886

AN - SCOPUS:0142039017

VL - 550

SP - 57

EP - 63

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 1-3

ER -

Access to Document

10.1016/S0014-5793(03)00817-2