Role of PDZK1 in lipid metabolism and atherosclerosis

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Adaptor, anchoring and scaffolding proteins play an important role in signal transduction. In many situations, the interaction between signaling proteins is mediated by small amino acid sequences binding to specific protein domains, such as src homology (SH), pTyr-binding (PTB) or PDZ domains. These interactions are responsible for determining location, function and activity of receptor and transporter proteins among others. A few years ago, we isolated a novel protein that we named PDZK1. PDZK1 contains four PDZ protein-interaction domains and interacts with the carboxy-terminal portion of a number of membrane associated proteins including the high density lipoprotein (HDL) scavenger receptor SR-BI and several ion channels, one of them involved in multidrug resistance. As a result, PDZK1 is likely to play an important role in biological processes as diverse as lipid metabolism and cardiovascular disease, ion channel organization and multidrug resistance. We generated a PDZK1 knockout mouse which is characterized by increased plasma cholesterol levels and markedly reduced expression of SR-BI in the liver, resulting in impaired "reverse cholesterol transport". PDZK1 joins ARH (the product of the defective gene in autosomal recessive hypercholesterolemia) in what is likely to be a growing family of cytoplasmic adaptor proteins that control the tissue specific activity of cell surface receptors. More recently, we have determined, using a transgenic mouse model in which truncated forms of PDZK1 were overexpressed in the liver of PDZK1 KO mice, that all four PDZ domains of PDZK1 are necessary for normal SR-BI expression, localization and function, although the precise role of individual domains, PDZ2 and PDZ4 in particular, remains unknown. We have determined that SR-BI is able to interact with another site within PDZK1, in addition to the known interaction with PDZ1. In addition, we have solved the crystal structure of the first domain of PDZK1 and its interaction with SR-BI. We have also determined that PDZK1 is atheroprotective, suggesting that PDZK1 may be an attractive target for therapies in cardiovascular diseases. Our objectives are now to study the molecular organization of the PDZK1/SR-BI protein cluster. We will define the functions of the second and fourth PDZ domains (PDZ2 and PDZ4) of PDZK1 and identify their binding partners, define the complete structure of PDZK1 by X-ray crystallography to obtain a three dimensional structure of the molecule and understand how PDZK1 interacts with its binding partners. We will correlate the results obtained in vitro using a transgenic mouse model, where altered (mutated/truncated) PDZK1 transgenes will be expressed in the liver of PDZK1 KO mice. Such studies may provide the necessary background for the development of new therapies promoting reverse cholesterol transport to prevent the formation of atherosclerotic lesions and myocardial infarction.