Western blot analysis of HIF1 Alpha in HeLa cell lysates using a 1:500 dilution of SMC-184.

Hypoxia-inducible factor 1 (HIF1) is a heterodimeric transcription factor that plays a critical role in the cellular response of hypoxia (1). The HIF1 complex consists of two subunits, HIF1-Alpha and HIF1-Beta, which are basic helix-loop-helix proteins of the PAS family (2). HIF1 regulates the transcription of a broad range of genes that facilitate responses to the hypoxic environment, including genes regulating angiogenesis, erythropoiesis, cell cycle, metabolism and apoptosis. The widely expressed HIF-1α is typically degraded rapidly in normoxic cells by the ubiquitin/proteasomal pathway. Under normoxic conditions, HIF-1α is proline hydroxylated leading to a conformational change that promotes binding to the von Hippel Lindau protein (VLH) E3 ligase complex; ubiquitination and proteasomal degradation follows (3, 4). Both hypoxic conditions and chemical hydroxylase inhibitors (such as desferrioxamine and cobalt) inhibit HIF-1α degradation and lead to its stabilization. In addition, HIF-1α can be induced in an oxygen-independent manner by various cytokines through the PI3K-AKT-mTOR pathway (5-7).

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1. Surazynski, A., Miltyk, W., Prokop, I. and Palka, J. (2013). The effect of estrogen on prolidase-dependent regulation of HIF-1α expression in breast cancer cells. Mol and Cel Biochem. doi:10.1007/s11010-013-1623-9 (includes protocol for mortalin/Grp75 ELISA).