Abstract
Publisher Summary This chapter focuses on oxygen sensing during the analysis of von Hippel–Lindau (VHL) hereditary cancer syndrome and the implications of oxygen sensing. VHL disease is a hereditary cancer syndrome caused by germline mutations that inactivate the VHL tumor suppressor gene that resides on chromosome 3p25. The cardinal features of this disorder are blood vessel tumors (hemangioblastomas) of the central nervous system (CNS) and retina, clear cell renal carcinomas, and pheochromocytomas. Tumor development in VHL disease is linked to somatic inactivation of the remaining wild-type VHL allele, thus depriving a susceptible cell of the wild-type VHL gene product (pVHL).The chapter discusses the expression and purification of GST–VBC complex and VBC complex, production of pure VBC complex and GST–VBC for pull down, far western blot analysis with VBC complex, i n vitro ubiquitination assay, and prolyl hydroxylation protocols
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