| Purification and Quality Control | The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified VHL is greater than 90% homogeneous based on SDS-PAGE analysis. | | Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. 30-100 units (ng) are required for an in vitro transcription assay and protein-protein interaction assay. | | Applications | Purified VHL protein has been used for in vitro transcriptional activation and protein-protein interaction assays. | | Formulation and Storage | The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles. | | Synonym | HRCA1; RCA1; VHL1 | | Protein Sequence | MPRRAENWDE AEVGAEEAGV EEYGPEEDGG EESGAEESGP EESGPEELGA EEEMEAGRPR
PVLRSVNSRE PSQVIFCNRS PRVVLPVWLN FDGEPQPYPT LPPGTGRRIH SYRGHLWLFR
DAGTHDGLLV NQTELFVPSL NVDGQPIFAN ITLPVYTLKE RCLQVVRSLV KPENYRRLDI
VRSLYEDLED HPNVQKDLER LTQERIAHQR MGD | | Background | von Hippel-Lindau (VHL) disease is a hereditary cancer with a predilection for the central nervous system and retina (1). The von Hippel-Lindau tumor suppressor gene is mutated in families with von Hippel-Lindau disease and encodes a protein (VHL) of 213 amino acids with an acidic pentapeptide motif in the N-terminus (2-4). Mutations in the VHL gene result in constitutive expression of many hypoxi-induced genes, at least in part because of increases in the cellular level of hypoxia-inducible transcription factor HIF-1a (5). VHL protein binds to elongin B, elongin C, and Cul2 to form a stable complex that targets hypoxia inducibal factors (HIFs) for degradation and transcriptional regulation (6,7). In addition, VHL protein has also been shown to interact with specific protein kinase C isoforms (8), histone deacetylases and HIF-1 inhibitor (FIH-1) (9). | | References | 1. Singh, AD. et al., (2001) Survey of Ophthalmology 46, 117-134
2. Latif, F. et al., (1993) Science 260, 1317-1320
3. Linehan, WM. et al., (1995) J. Am. Med. Assoc. 273, 564-570
4. Duan, DR. et al., (1995) Proc. Natl. Acad. Sci. USA 92, 6459-6463
5. Kamura, T. et al., (2000) Proc. Natl. Acad. Sci. USA 97, 10430-10435
6. Duan, DR. et al., (1995) Science 269, 1402-1406
7. Iwai, K. et al., (1999) Proc. Natl. Acad. Sci. USA 96, 12436-12441
8. Pal, S. et al., (1997) J Biol. Chem. 272, 27509-27512
9. Mahon, PC. et al., (2001) Genes & Dev. 15, 2675-2686 |
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