boltzmann energy-based image analysis demonstrates that extracellular domain size differences explain protein segregation at immune synapses玻耳兹曼能源图像分析表明,细胞外域大小差异解释蛋白质隔离在免疫突触.pdfVIP

Boltzmann Energy-based Image Analysis Demonstrates that Extracellular Domain Size Differences Explain Protein Segregation at Immune Synapses 1 ¨ 2 1 3 Nigel J. Burroughs *, Karsten Kohler , Vladimir Miloserdov , Michael L. Dustin , P. Anton van der Merwe4, Daniel M. Davis5 1 Systems Biology Centre, University of Warwick, Coventry, United Kingdom, 2 Cambridge Institute for Medical Research, Addenbrookes Hospital, Cambridge, United Kingdom, 3 Skirball Institute of Biomolecular Medicine, New York University, New York, New York, United States of America, 4 Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom, 5 Division of Cell and Molecular Biology, Imperial College London, London, United Kingdom Abstract Immune synapses formed by T and NK cells both show segregation of the integrin ICAM1 from other proteins such as CD2 (T cell) or KIR (NK cell). However, the mechanism by which these proteins segregate remains unclear; one key hypothesis is a redistribution based on protein size. Simulations of this mechanism qualitatively reproduce observed segregation patterns, but only in certain parameter regimes. Verifying that these parameter constraints in fact hold has not been possible to date, this requiring a quantitative coupling of theory to experimental data. Here, we address this challenge, developing a new methodology for analysing and quantifying image data and its integration with biophysical models. Specifically we fit a binding kinetics model to 2 colour fluorescence data for cytoskeleton independent synapses (2 and 3D) and test whether the observed inverse correlation between fluorophores conforms to size dependent exclusion, and further, whether patterned states are predicted when model p