ALL RAW DATA FOR : Pathological mutations differentially affect the self-assembly and polymerisation of the the immune system signaling adaptor molecule MyD88

2018-11-08T05:39:16Z (GMT) by Interacteam UNSW
The fluorescence time-traces obtained for the full-length MYD88, mutants of MYD88 and the separate domains of MyD88 from single molecule fluorescence spectroscopy. For TIR domain alone the fluctuations of intensity around the average value are limited, for the death domain, small bursts of intensities can be detected. These fluorescence peaks correspond to entries of single protein complexes, increasing the local number of proteins for a brief period of time. The amplitude and duration of the deviations from average are linked to the number of proteins co-diffusing in a single complex and the physical size of the diffusion complex. For full-length MyD88, we observe extremely bright and long-diffusing bursts of fluorescence. The simplest analysis to quantify the presence of protein complexes is to calculate the average brightness of the diffusing species, calculated from the measured intensity values (I) as B=〖(Standard deviation (I))〗^2/(average (I)). FCS data in solution. Values are from repeated dilution experiments with the various protein concentrations and corresponding brightness values obtained plotted. NS >0.9999, ***P=0.0001, ****P<0.0001