Dual applications of a c-Myc antibody in mitochondrial research

Thu, 02/09/2017 - 08:10

c-Myc, a proto-oncogene, has documented involvement in cellular differentiation, cell growth, cell death and tumor formation.  Target genes of the Myc family include those that participate in cell survival, translation, transcription, metabolism and more.  On a more specific level, c-Myc is a transcription factor that can both activate and repress its target protein by way of DNA modifications.  This allows for the use of a c-Myc antibody in two manners; it can be used to monitor the actual c-Myc protein expression levels, or, it can be used as a probe against a c-Myc protein tagged to another protein of interest. These interactions can be exploited when trying to detect proteins that c-myc interacts with if antibodies against the protein are not available. The structure of c-Myc is what allows for its duality, where its basic-helix-loop-helix-leucine zipper c-term domain can mediate DNA specific binding.  The following articles explain the use of the c-Myc antibody as both a probe for a myc-tagged protein and also as a protein of interest in its natural form.

c-myc antibody

c-Myc Antibody (9E10) [NB600-302] - Analysis of c-myc in Jurkat cell lysates using NB600-302.

First, Jang et al used a c-Myc antibody to show that mitochondrial function provides instructive signaling for activated-induced B-cell fates.  Due to the fact that the analysis of metabolic pathways relies on lineage cellular differentiation, two cellular populations were established – P1 and P2 cell lines.  P1 had a more uniform population of B cells that demonstrated increased mitochondrial mass and membrane potential, and P2 cells had further increased mitochondrial mass and membrane potential and further differentiation.  A c-Myc antibody was used during the characterization of these two cell populations, alongside Pax5, Bcl6 and AID antibodies, as these proteins had been reported to promote class-switch recombination.  Specifically, the c-Myc antibody was used in western blot.

Next, Pekkurnaz et al used a c-Myc antibody as a probe against a myc-hMilton construct in their research of glucose regulated mitochondrial motility via Milton modification.  Milton is a mitochondrial motor-adaptor protein that is a required substrate for the enzyme O-GlcNAc Transferase (OGT) that is particularly import to mitochondrial dynamics in neurons.  To first test the theory that OGT disrupts mitochondrial motility via dissociation of the Milton/KHC/Miro complex, hMilton1 was immunoprecipitated from HEK293T cells with or without enhanced OGT expression.  The c-Myc antibody identified a band in these HEK293T precipitates that concluded alteration sites on Milton are governed by a balance of OGT.  The c-Myc antibody was also used to confirm that OGT dependent mitochondrial motility arrest requires Milton through a co-immunostaining protocol in HEK293T cells.  Neurons were co-stained for Mito-DsRed and the c-Myc antibody, and were found to have an extremely high correlation of co-staining.  Overall, these results in addition to further experiments suggest that the OGT pathway is another important signal cascade that affects mitochondrial function. 

View all c-Myc antibodies for your research.

  1. Dang CV. MYC on the path to cancer.[PMID: 22464321]
  2. Pekkurnaz G, Trinidad JC, Wang X, Kong D, Schwarz TL. Glucose regulates mitochondrial motility via Milton modification by O-GlcNAc transferase. [PMID: 24995978]
  3. Hoffman B, Liebermann DA. Apoptotic signaling by c-MYC. [PMID: 18955973]
  4. Li F, Wang Y, Zeller KI, Potter JJ, Wonsey DR, O'Donnell KA, Kim JW, Yustein JT, Lee LA, Dang CV. Myc stimulates nuclearly encoded mitochondrial genes and mitochondrial biogenesis. [PMID: 15988031]

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