title: Casein Kinase 2 Is a Novel Regulator of the Human Organic Anion Transporting Polypeptide 1A2 (OATP1A2) Trafficking.
abstract: Human organic anion transporting polypeptides (OATPs) mediate the influx of many important drugs into cells. Casein kinase 2 (CK2) is a critical protein kinase that phosphorylates >300 protein substrates and is dysregulated in a number of disease states. Among the CK2 substrates are several transporters, although whether this includes human OATPs has not been evaluated. The current study was undertaken to evaluate the regulation of human OATP1A2 by CK2. HEK-239T cells in which OATP1A2 was overexpressed were treated with CK2 specific inhibitors or transfected with CK2 specific siRNA, and the activity, expression, and subcellular trafficking of OATP1A2 was evaluated. CK2 inhibition decreased the uptake of the prototypic OATP1A2 substrate estrone-3-sulfate (E3S). Kinetic studies revealed that this was due to a decrease in the maximum velocity (Vmax) of E3S uptake, while the Michaelis constant was unchanged. The cell surface expression, but not the total cellular expression of OATP1A2, was impaired by CK2 inhibition and knockdown of the catalytic α-subunits of CK2. CK2 inhibition decreased the internalization of OATP1A2 via a clathrin-dependent pathway, decreased OATP1A2 recycling, and likely impaired OATP1A2 targeting to the cell surface. Consistent with these findings, CK2 inhibition also disrupted the colocalization of OATP1A2 and Rab GTPase (Rab)4-, Rab8-, and Rab9-positive endosomal and secretory vesicles. Taken together, CK2 has emerged as a novel regulator of the subcellular trafficking and stability of OATP1A2. Because OATP1A2 transports many molecules of physiological and pharmacological importance, the present data may inform drug selection in patients with diseases in which CK2 and OATP1A2 are dysregulated.
authors.affiliation: Faculty of Pharmacy, The University of Sydney , Sydney, New South Wales 2006, Australia.
authors.forename: Ting
authors.initials: T
authors.lastname: Chan
authors.affiliation: Faculty of Pharmacy, The University of Sydney , Sydney, New South Wales 2006, Australia.
authors.forename: Florence Shin Gee
authors.initials: FS
authors.lastname: Cheung
authors.affiliation: Alkali Soil Natural Environmental Science Center, Northeast Forestry University/Key Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field, Ministry of Education , Harbin, 150040, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Zheng
authors.affiliation: Faculty of Pharmacy, The University of Sydney , Sydney, New South Wales 2006, Australia.
authors.forename: Xiaoxi
authors.initials: X
authors.lastname: Lu
authors.affiliation: Retinal Therapeutics Research Group, Save Sight Institute, The University of Sydney , Sydney, New South Wales 2000, Australia.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Faculty of Pharmacy, The University of Sydney , Sydney, New South Wales 2006, Australia.
authors.forename: Thomas
authors.initials: T
authors.lastname: Grewal
authors.affiliation: Discipline of Pharmacology, School of Medical Sciences, The University of Sydney , Sydney, New South Wales 2006, Australia.
authors.forename: Michael
authors.initials: M
authors.lastname: Murray
authors.affiliation: Faculty of Pharmacy, The University of Sydney , Sydney, New South Wales 2006, Australia.
authors.forename: Fanfan
authors.initials: F
authors.lastname: Zhou
mesh_terms: D047390:Casein Kinase II / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D020868:Gene Silencing / Q000502:physiology; D006801:Humans; D015151:Immunoblotting; D027361:Organic Anion Transporters / Q000378:metabolism*; D055550:Protein Stability; D021381:Protein Transport / Q000502:physiology
chemical_list: D027361:Organic Anion Transporters; D047390:Casein Kinase II
keywords: casein kinase 2; human organic anion transporting polypeptide 1A2; membrane trafficking; molecular regulation; protein stability