Changes in copper concentrations affect the protein levels but not the mRNA levels of copper chaperones in human umbilical vein endothelial cells

Daoyin Dong, Xinhua Xu, Wen Yin, Yujian Kang

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Copper chaperones are critical regulators of intracellular copper metabolism and distribution. The present study was undertaken to investigate the effects of changes in copper concentrations on the abundance of copper chaperones. Human umbilical vein endothelial cells (HUVECs) were treated with siRNA targeting copper transporter 1 (CTR1) or tetraethylenepentamine (TEPA) to decrease, or with copper sulfide to increase, intracellular copper concentrations, assayed using an atomic absorption spectrophotometer. Western blot analyses showed that CTR1 silencing or TEPA treatment increased the protein levels of copper chaperone ATOX1 and copper chaperone for superoxide dismutase 1 (CCS-1), but decreased copper chaperone for cytochrome c oxidase (COX17). In contrast, copper supplementation decreased the protein levels of ATOX1 and CCS-1 and increased COX17. Real-time RT-PCR analyses found no difference in the mRNA levels of the copper chaperones examined under the conditions defined above. This study thus demonstrated that changes in copper concentrations alter the protein levels, but not the mRNA levels, of copper chaperones, suggesting a role of copper in the post-translational modification of these proteins.

Original languageEnglish (US)
Pages (from-to)554-559
Number of pages6
JournalMetallomics
Volume6
Issue number3
DOIs
StatePublished - Mar 1 2014
Externally publishedYes

Fingerprint

Endothelial cells
Human Umbilical Vein Endothelial Cells
Copper
Proteins
Messenger RNA
Superoxide Dismutase
Sulfides
Electron Transport Complex IV
Post Translational Protein Processing
Spectrophotometers
Small Interfering RNA
Metabolism
Real-Time Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Chemistry (miscellaneous)
  • Biophysics
  • Biomaterials
  • Biochemistry
  • Metals and Alloys

Cite this

Changes in copper concentrations affect the protein levels but not the mRNA levels of copper chaperones in human umbilical vein endothelial cells. / Dong, Daoyin; Xu, Xinhua; Yin, Wen; Kang, Yujian.

In: Metallomics, Vol. 6, No. 3, 01.03.2014, p. 554-559.

Research output: Contribution to journalArticle

@article{40216239b45f438d97a1f6d640820cf3,
title = "Changes in copper concentrations affect the protein levels but not the mRNA levels of copper chaperones in human umbilical vein endothelial cells",
abstract = "Copper chaperones are critical regulators of intracellular copper metabolism and distribution. The present study was undertaken to investigate the effects of changes in copper concentrations on the abundance of copper chaperones. Human umbilical vein endothelial cells (HUVECs) were treated with siRNA targeting copper transporter 1 (CTR1) or tetraethylenepentamine (TEPA) to decrease, or with copper sulfide to increase, intracellular copper concentrations, assayed using an atomic absorption spectrophotometer. Western blot analyses showed that CTR1 silencing or TEPA treatment increased the protein levels of copper chaperone ATOX1 and copper chaperone for superoxide dismutase 1 (CCS-1), but decreased copper chaperone for cytochrome c oxidase (COX17). In contrast, copper supplementation decreased the protein levels of ATOX1 and CCS-1 and increased COX17. Real-time RT-PCR analyses found no difference in the mRNA levels of the copper chaperones examined under the conditions defined above. This study thus demonstrated that changes in copper concentrations alter the protein levels, but not the mRNA levels, of copper chaperones, suggesting a role of copper in the post-translational modification of these proteins.",
author = "Daoyin Dong and Xinhua Xu and Wen Yin and Yujian Kang",
year = "2014",
month = "3",
day = "1",
doi = "10.1039/c3mt00138e",
language = "English (US)",
volume = "6",
pages = "554--559",
journal = "Metallomics",
issn = "1756-5901",
publisher = "Royal Society of Chemistry",
number = "3",

}

TY - JOUR

T1 - Changes in copper concentrations affect the protein levels but not the mRNA levels of copper chaperones in human umbilical vein endothelial cells

AU - Dong, Daoyin

AU - Xu, Xinhua

AU - Yin, Wen

AU - Kang, Yujian

PY - 2014/3/1

Y1 - 2014/3/1

N2 - Copper chaperones are critical regulators of intracellular copper metabolism and distribution. The present study was undertaken to investigate the effects of changes in copper concentrations on the abundance of copper chaperones. Human umbilical vein endothelial cells (HUVECs) were treated with siRNA targeting copper transporter 1 (CTR1) or tetraethylenepentamine (TEPA) to decrease, or with copper sulfide to increase, intracellular copper concentrations, assayed using an atomic absorption spectrophotometer. Western blot analyses showed that CTR1 silencing or TEPA treatment increased the protein levels of copper chaperone ATOX1 and copper chaperone for superoxide dismutase 1 (CCS-1), but decreased copper chaperone for cytochrome c oxidase (COX17). In contrast, copper supplementation decreased the protein levels of ATOX1 and CCS-1 and increased COX17. Real-time RT-PCR analyses found no difference in the mRNA levels of the copper chaperones examined under the conditions defined above. This study thus demonstrated that changes in copper concentrations alter the protein levels, but not the mRNA levels, of copper chaperones, suggesting a role of copper in the post-translational modification of these proteins.

AB - Copper chaperones are critical regulators of intracellular copper metabolism and distribution. The present study was undertaken to investigate the effects of changes in copper concentrations on the abundance of copper chaperones. Human umbilical vein endothelial cells (HUVECs) were treated with siRNA targeting copper transporter 1 (CTR1) or tetraethylenepentamine (TEPA) to decrease, or with copper sulfide to increase, intracellular copper concentrations, assayed using an atomic absorption spectrophotometer. Western blot analyses showed that CTR1 silencing or TEPA treatment increased the protein levels of copper chaperone ATOX1 and copper chaperone for superoxide dismutase 1 (CCS-1), but decreased copper chaperone for cytochrome c oxidase (COX17). In contrast, copper supplementation decreased the protein levels of ATOX1 and CCS-1 and increased COX17. Real-time RT-PCR analyses found no difference in the mRNA levels of the copper chaperones examined under the conditions defined above. This study thus demonstrated that changes in copper concentrations alter the protein levels, but not the mRNA levels, of copper chaperones, suggesting a role of copper in the post-translational modification of these proteins.

UR - http://www.scopus.com/inward/record.url?scp=84896818142&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84896818142&partnerID=8YFLogxK

U2 - 10.1039/c3mt00138e

DO - 10.1039/c3mt00138e

M3 - Article

VL - 6

SP - 554

EP - 559

JO - Metallomics

JF - Metallomics

SN - 1756-5901

IS - 3

ER -