Multiple oxidative stress-response members of the Adapt78 family

Henry J. Michtalik, Ananth V. Narayan, Nishant Bhatt, H. Y. Lin, Michael T. Mulligan, Shen Li Zhang, Dana R. Crawford

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Adapt78 is an oxidative and calcium stress-response gene. Its protein product is a potent natural inhibitor of the intracellular calcium signaling protein calcineurin. Much of what is known about Adapt78 protein is based on cell-transfection studies. Toward understanding natural endogenous Adapt78, we used an antibody raised against cellular Adapt78 and recently determined that endogenous Adapt78 protein, like its mRNA, is oxidative and calcium stress responsive. Here we report the identification of a second endogenous form of this protein family of 41 kDa. Subcellular fractionation of human HeLa cells revealed that in contrast to results of previous transfection studies, most endogenous Adapt78, characterized as 29 and 41 kDa electrophoretic doublets, resides in the cellular cytosol. The 41 kDa form of Adapt78 was abundant and found to exhibit many characteristics in common with the previously reported oxidative stress-responsive 29 kDa form, including hypo- and hyperphosphorylation variants, rapid loss of the hypophosphorylated form following oxidative stress, response to various kinase and phosphatase inhibitors, and localization. However, it also exhibited some unique characteristics, most notably the lack of calcium inducibility. Finally, the 29 kDa form exhibited a much shorter half-life and strong stabilization following oxidant exposure compared with the 41 kDa Adapt78 form. These data reveal the presence of a novel oxidative stress-responsive 41 kDa Adapt78 species, lend further insight into the Adapt78 family of proteins and their distribution, and challenge previous conclusions obtained using transfection protocols.

Original languageEnglish
Pages (from-to)454-462
Number of pages9
JournalFree Radical Biology and Medicine
Volume37
Issue number4
DOIs
Publication statusPublished - Aug 15 2004
Externally publishedYes

Fingerprint

Oxidative stress
Oxidative Stress
Transfection
Calcium
Proteins
Intracellular Signaling Peptides and Proteins
Calcium Signaling
Calcineurin
HeLa Cells
Phosphoric Monoester Hydrolases
Oxidants
Cytosol
Half-Life
Fractionation
Phosphotransferases
Stabilization
Genes
Messenger RNA
Antibodies

Keywords

  • DMEM
  • Dulbecco's minimal essential medium
  • FCS
  • fetal calf serum
  • HP
  • hydrogen peroxide
  • HyperP
  • hyperphosphorylated
  • HypoP
  • hypophosphorylated
  • lambda phosphatase
  • LP
  • Nitrosative stress
  • NonP
  • nonphosphorylated
  • Oxidative stress
  • PAO
  • peroxynitrite
  • phenylarsine
  • PN

ASJC Scopus subject areas

  • Medicine(all)
  • Toxicology
  • Clinical Biochemistry

Cite this

Michtalik, H. J., Narayan, A. V., Bhatt, N., Lin, H. Y., Mulligan, M. T., Zhang, S. L., & Crawford, D. R. (2004). Multiple oxidative stress-response members of the Adapt78 family. Free Radical Biology and Medicine, 37(4), 454-462. https://doi.org/10.1016/j.freeradbiomed.2004.05.014

Multiple oxidative stress-response members of the Adapt78 family. / Michtalik, Henry J.; Narayan, Ananth V.; Bhatt, Nishant; Lin, H. Y.; Mulligan, Michael T.; Zhang, Shen Li; Crawford, Dana R.

In: Free Radical Biology and Medicine, Vol. 37, No. 4, 15.08.2004, p. 454-462.

Research output: Contribution to journalArticle

Michtalik, HJ, Narayan, AV, Bhatt, N, Lin, HY, Mulligan, MT, Zhang, SL & Crawford, DR 2004, 'Multiple oxidative stress-response members of the Adapt78 family', Free Radical Biology and Medicine, vol. 37, no. 4, pp. 454-462. https://doi.org/10.1016/j.freeradbiomed.2004.05.014
Michtalik, Henry J. ; Narayan, Ananth V. ; Bhatt, Nishant ; Lin, H. Y. ; Mulligan, Michael T. ; Zhang, Shen Li ; Crawford, Dana R. / Multiple oxidative stress-response members of the Adapt78 family. In: Free Radical Biology and Medicine. 2004 ; Vol. 37, No. 4. pp. 454-462.
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