Study of transport of protein molecules across cell membranes

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Aneta Hornová Supervisor: PharmDr. Lucie Hyršová, Ph.D. Title of diploma thesis: Study of transport of protein molecules across cell membranes Kidney transporters are found mainly in the proximal tubules, they participate in the secretion and reabsorption of endogenous and exogenous substances. Plenty of kidney transporters have been identified, however this diploma thesis is focused on megalin. Specifically, we dealed with determination of transport of peptides via this renal transporter in HK-2 and JEG-3 cell lines. Megalin is a glycoprotein whose main function is tubular reabsorption of albumin filtered in glomerules and other high molecular weight substances. Another endogenous substrates of megalin are e.g. transcoblamine - vitamin B12, insulin or light chains of immunoglobulins. It is also able to transport some drugs like aminoglycoside and polypeptide antibiotics. Within this work, we investigated the influence of reduced expression of megalin on the transport of albumin and anti-VEGFR2 aptamer. Reduction of expression was achieved by transfection of cells with siRNA against the LRP2 gene. Lower expression of LRP2 led to reduction of transport of albumin and anti-VEGFR2 aptamer into..