LMP402 - Lecture 8.doc

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Laboratory Medicine and Pathobiology
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LMP402Fall 2013Lecture 8 TLR4 LPS LBP mice susceptible to infection MD2mice susceptible to infection WHEN infect with Salmonella and count number of surviving mice TLR4 surface expression requires MD3fluorescent labelling of TLR4Alanine scan of MD2where every amino acid mutated to alanine then measure responsiveness to LPS Graph LPS responsiveness versus cell surface expressionshould expect a rising slanted line with slope of 1 expected if MD2 only has functions of surface expression However have mutants above and below this expected line suggesting MD2 mutations may diminish binding to ligands or impair signalling Crystallography determined direct binding of LPS to MD2MD2 function in cell surface TLR4 localization and LPS Response TLR4 forms horseshoe shape with MD2 in horseshoe LPS forms bridge that lins the 2 protein complexes Number of acyl chains important for whether LPS is detected or not by TLR4Not enough chains wont form bridge no detection Bacteria can modulate LPS structurePseudomonas aeuringinosa5 chains in lab strain 6 chains in human CF patient Human cells are defective at detecting pentaacylated LPS Human cells and labadapte
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