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 journalArticle

26 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 languageEnglish
Pages (from-to)57-63
Number of pages7
JournalFEBS Letters
Volume550
Issue number1-3
DOIs
Publication statusPublished - Aug 28 2003
Externally publishedYes

Fingerprint

Calcium-Calmodulin-Dependent Protein Kinases
HeLa Cells
Phosphotransferases
Chemical activation
Phosphorylation
Ionomycin
Protein Kinases
Protein Isoforms
Mutation

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Identification and characterization of novel components of a Ca 2+/calmodulin-dependent protein kinase cascade in HeLa cells. / Ishikawa, Yumi; Tokumitsu, Hiroshi; Inuzuka, Hiroyuki; Murata-Hori, Maki; Hosoya, Hiroshi; Kobayashi, Ryoji.

In: FEBS Letters, Vol. 550, No. 1-3, 28.08.2003, p. 57-63.

Research output: Contribution to journalArticle

Ishikawa, Yumi ; Tokumitsu, Hiroshi ; Inuzuka, Hiroyuki ; Murata-Hori, Maki ; Hosoya, Hiroshi ; Kobayashi, Ryoji. / Identification and characterization of novel components of a Ca 2+/calmodulin-dependent protein kinase cascade in HeLa cells. In: FEBS Letters. 2003 ; Vol. 550, No. 1-3. pp. 57-63.
@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 = "8",
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 -