[25] Identification of in vivo phosphorylation sites for basic-directed kinases in murine mdr1b P-glycoprotein by combination of mass spectrometry and site-directed mutagenesis

摘要: Publisher Summary This chapter discusses the identification of in vivo phosphorylation sites for basic-directed kinases murine mdr1b P-glycoprotein by a combination mass spectrometry and site-directed mutagenesis. Reversible phosphorylation/dephosphorylation proteins on serine, threonine, tyrosine residues constitutes an important mechanism regulating activity eukaryotic cells. It is known that multidrug resistant (MDR) transporter, P-glycoprotein, phosphorylated but relevance this posttranslational modification to transporter function poorly understood. The approach described investigates physiologic map then mutate identified sites, singly or combination, non-phosphorylated residues. creation stably transfected cell lines carrying desired mutations allows dissection role function.