LMP299Y1 Lecture Notes - Lecture 8: Crystallography, Alanine, Acylation

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Lmp402 - fall 2013 - lecture 8: tlr4, lps. When infect with salmonella, and count number of surviving mice. Tlr4 surface expression requires md3 - fluorescent labelling of tlr4. Alanine scan of md2 - where every amino acid mutated to alanine, then measure responsiveness to lps. Graph lps responsiveness versus cell surface expression - should 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 md2. Md2 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 tlr4. Not enough chains, won"t form bridge, no detection.

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