The Tumor-sensitive Calmodulin-like Protein Is a Specific Light Chain of Human Unconventional Myosin X

Michael S. Rogers, Emanuel E. Strehler

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Human calmodulin-like protein (CLP) is an epithelial-specific Ca 2+-binding protein whose expression is strongly down-regulated in cancers. Like calmodulin, CLP is thought to regulate cellular processes via Ca 2+-dependent interactions with specific target proteins. Using gel overlays, we identified a ∼-210-kDa protein binding specifically and in a Ca 2+-dependent manner to CLP, but not to calmodulin. Yeast two-hybrid screening yielded a CLP-interacting clone encoding the three light chain binding IQ motifs of human "unconventional" myosin X. Pull-down experiments showed CLP binding to the IQ domain to be direct and Ca 2+-dependent. CLP interacted strongly with IQ motif 3 (Kd ∼0.5 nM) as determined by surface plasmon resonance. Epitope-tagged myosin X was localized preferentially at the cell periphery in MCF-7 cells, and CLP colocalized with myosin X in these cells. Myosin X was able to coprecipitate CLP and, to a lesser extent, calmodulin from transfected COS-1 cells, indicating that CLP is a specific light chain of myosin X in vivo. Because unconventional myosins participate in cellular processes ranging from membrane trafficking to signaling and cell motility, myosin X is an attractive CLP target. Altered myosin X regulation in (tumor) cells lacking CLP may have as yet unknown consequences for cell growth and differentiation.

Original languageEnglish (US)
Pages (from-to)12182-12189
Number of pages8
JournalJournal of Biological Chemistry
Volume276
Issue number15
DOIs
StatePublished - Apr 13 2001

Fingerprint

Calmodulin
Myosins
Tumors
Light
Neoplasms
Proteins
Calmodulin-Binding Proteins
Myosin Light Chains
Surface Plasmon Resonance
COS Cells
MCF-7 Cells
Cell growth
Surface plasmon resonance
Protein Binding
Cell Movement
Yeast
Epitopes
Cell Differentiation
Clone Cells
Yeasts

ASJC Scopus subject areas

  • Biochemistry

Cite this

The Tumor-sensitive Calmodulin-like Protein Is a Specific Light Chain of Human Unconventional Myosin X. / Rogers, Michael S.; Strehler, Emanuel E.

In: Journal of Biological Chemistry, Vol. 276, No. 15, 13.04.2001, p. 12182-12189.

Research output: Contribution to journalArticle

Rogers, Michael S. ; Strehler, Emanuel E. / The Tumor-sensitive Calmodulin-like Protein Is a Specific Light Chain of Human Unconventional Myosin X. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 15. pp. 12182-12189.
@article{bc9fd1cbce27491a9694a27b0fa70760,
title = "The Tumor-sensitive Calmodulin-like Protein Is a Specific Light Chain of Human Unconventional Myosin X",
abstract = "Human calmodulin-like protein (CLP) is an epithelial-specific Ca 2+-binding protein whose expression is strongly down-regulated in cancers. Like calmodulin, CLP is thought to regulate cellular processes via Ca 2+-dependent interactions with specific target proteins. Using gel overlays, we identified a ∼-210-kDa protein binding specifically and in a Ca 2+-dependent manner to CLP, but not to calmodulin. Yeast two-hybrid screening yielded a CLP-interacting clone encoding the three light chain binding IQ motifs of human {"}unconventional{"} myosin X. Pull-down experiments showed CLP binding to the IQ domain to be direct and Ca 2+-dependent. CLP interacted strongly with IQ motif 3 (Kd ∼0.5 nM) as determined by surface plasmon resonance. Epitope-tagged myosin X was localized preferentially at the cell periphery in MCF-7 cells, and CLP colocalized with myosin X in these cells. Myosin X was able to coprecipitate CLP and, to a lesser extent, calmodulin from transfected COS-1 cells, indicating that CLP is a specific light chain of myosin X in vivo. Because unconventional myosins participate in cellular processes ranging from membrane trafficking to signaling and cell motility, myosin X is an attractive CLP target. Altered myosin X regulation in (tumor) cells lacking CLP may have as yet unknown consequences for cell growth and differentiation.",
author = "Rogers, {Michael S.} and Strehler, {Emanuel E.}",
year = "2001",
month = "4",
day = "13",
doi = "10.1074/jbc.M010056200",
language = "English (US)",
volume = "276",
pages = "12182--12189",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "15",

}

TY - JOUR

T1 - The Tumor-sensitive Calmodulin-like Protein Is a Specific Light Chain of Human Unconventional Myosin X

AU - Rogers, Michael S.

AU - Strehler, Emanuel E.

PY - 2001/4/13

Y1 - 2001/4/13

N2 - Human calmodulin-like protein (CLP) is an epithelial-specific Ca 2+-binding protein whose expression is strongly down-regulated in cancers. Like calmodulin, CLP is thought to regulate cellular processes via Ca 2+-dependent interactions with specific target proteins. Using gel overlays, we identified a ∼-210-kDa protein binding specifically and in a Ca 2+-dependent manner to CLP, but not to calmodulin. Yeast two-hybrid screening yielded a CLP-interacting clone encoding the three light chain binding IQ motifs of human "unconventional" myosin X. Pull-down experiments showed CLP binding to the IQ domain to be direct and Ca 2+-dependent. CLP interacted strongly with IQ motif 3 (Kd ∼0.5 nM) as determined by surface plasmon resonance. Epitope-tagged myosin X was localized preferentially at the cell periphery in MCF-7 cells, and CLP colocalized with myosin X in these cells. Myosin X was able to coprecipitate CLP and, to a lesser extent, calmodulin from transfected COS-1 cells, indicating that CLP is a specific light chain of myosin X in vivo. Because unconventional myosins participate in cellular processes ranging from membrane trafficking to signaling and cell motility, myosin X is an attractive CLP target. Altered myosin X regulation in (tumor) cells lacking CLP may have as yet unknown consequences for cell growth and differentiation.

AB - Human calmodulin-like protein (CLP) is an epithelial-specific Ca 2+-binding protein whose expression is strongly down-regulated in cancers. Like calmodulin, CLP is thought to regulate cellular processes via Ca 2+-dependent interactions with specific target proteins. Using gel overlays, we identified a ∼-210-kDa protein binding specifically and in a Ca 2+-dependent manner to CLP, but not to calmodulin. Yeast two-hybrid screening yielded a CLP-interacting clone encoding the three light chain binding IQ motifs of human "unconventional" myosin X. Pull-down experiments showed CLP binding to the IQ domain to be direct and Ca 2+-dependent. CLP interacted strongly with IQ motif 3 (Kd ∼0.5 nM) as determined by surface plasmon resonance. Epitope-tagged myosin X was localized preferentially at the cell periphery in MCF-7 cells, and CLP colocalized with myosin X in these cells. Myosin X was able to coprecipitate CLP and, to a lesser extent, calmodulin from transfected COS-1 cells, indicating that CLP is a specific light chain of myosin X in vivo. Because unconventional myosins participate in cellular processes ranging from membrane trafficking to signaling and cell motility, myosin X is an attractive CLP target. Altered myosin X regulation in (tumor) cells lacking CLP may have as yet unknown consequences for cell growth and differentiation.

UR - http://www.scopus.com/inward/record.url?scp=0035853701&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035853701&partnerID=8YFLogxK

U2 - 10.1074/jbc.M010056200

DO - 10.1074/jbc.M010056200

M3 - Article

C2 - 11278607

AN - SCOPUS:0035853701

VL - 276

SP - 12182

EP - 12189

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 15

ER -