, Volume 35, Issue 1, pp 167–175 | Cite as

Human Macrophage ATP7A is Localized in the trans-Golgi Apparatus, Controls Intracellular Copper Levels, and Mediates Macrophage Responses to Dermal Wounds

  • Ha Won Kim
  • Qilin Chan
  • Scott E. Afton
  • Joseph A. Caruso
  • Barry Lai
  • Neal L. Weintraub
  • Zhenyu QinEmail author


The copper transporter ATP7A has attracted significant attention since the discovery of its gene mutation leading to human Menkes disease. We previously reported that ATP7A is highly expressed in the human vasculature and identified a novel vascular function of ATP7A in modulation of the expression and activity of extracellular superoxide dismutase. We recently identified that ATP7A expression in THP-1 cells (a monocyte/macrophage model cell line) plays a role in the oxidation of low density lipoproteins, indicating that it is necessary to further investigate its expression and function in monocytes/macrophages. In the current study, we demonstrated the protein and mRNA expression of ATP7A in human peripheral blood mononuclear cell (PBMC)-derived macrophages and alveolar macrophages. ATP7A was strongly co-localized with the trans-Golgi apparatus in PBMC-derived macrophages. Intracellular copper, detected by synchrotron X-ray fluorescence microscopy, was found to be distributed to the nucleus and cytoplasm in human THP-1 cells. To confirm the role of endogenous ATP7A in macrophage copper homeostasis, we performed inductively coupled plasma mass spectrometry in murine peritoneal macrophages, which showed markedly increased intracellular copper levels in macrophages isolated from ATP7A-deficient mice versus control mice. Moreover, the role of ATP7A in regulating macrophage responses to dermal wounds was studied by introduction of control and ATP7A-downregulated THP-1 cells into dermal wounds of nude mice. Infiltration of THP-1 cells into the wounded area (detected by expression of human macrophage markers MAC2 and CD68) was reduced in response to downregulation of ATP7A, hinting decreased macrophage accumulation subsequent to dermal wounds. In summary, alongside our previous studies, these findings indicate that human macrophage ATP7A is localized in the trans-Golgi apparatus, regulates intracellular copper levels, and mediates macrophage responses to a dermal wound.


macrophage copper ATP7A 


calcein AM

Calcein acetoxymethyl ester


Cytosolic phospholipase A




Differential interference contrast


Inductively coupled plasma mass spectrometry


Peripheral blood mononuclear cell


Phosphate-buffered saline


Platelet-derived growth factor




Synchrotron X-ray fluorescence


Vascular endothelial growth factor


Vascular smooth muscle cell



This work was supported by a National Scientist Development Grant (0835268N) from the American Heart Association and grants HL-076684 and HL-62984 from the National Institute of Health. Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract No. DE-AC02-06CH11357. We thank Birgit Ehmer and Chet Closson for assisting with the immunofluorescence microscopy and confocal microscopy, Dr. Oyebode Olakanmi for developing the protocol to isolate human PBMC-derived macrophages, and Dr. Dennis McGraw for providing human alveolar macrophages.


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Ha Won Kim
    • 1
  • Qilin Chan
    • 1
    • 2
  • Scott E. Afton
    • 1
    • 2
  • Joseph A. Caruso
    • 2
  • Barry Lai
    • 3
  • Neal L. Weintraub
    • 1
  • Zhenyu Qin
    • 1
    • 4
    Email author
  1. 1.Division of Cardiovascular Disease, Department of Internal MedicineUniversity of CincinnatiCincinnatiUSA
  2. 2.Department of Chemistry, College of Arts and SciencesUniversity of CincinnatiCincinnatiUSA
  3. 3.X-Ray Science DivisionArgonne National LaboratoryArgonneUSA
  4. 4.Division of Vascular Surgery, Department of SurgeryUniversity of Texas Health Science Center at San AntonioSan AntonioUSA

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