probing opioid signalosome on neuroblastoma原子力显微镜对阿片类signalosome对神经母细胞瘤课件.ppt

ＡＦＭ　Ｐｒｏｂｉｎｇ　Ｏｐｉｏｄ　Ｓｉｇｎａｌｏｓｏｍｅ　Ｃｏｍｐｌｅｘ ＡＦＭ　ｓｃｈｅｍａｔｉｃ Functionalization of AFM tips Detection of antibody-receptor recognition - SHSY5Y cell line stably expresses the MOPR tagged with T7. Force measurements by AFM Forces between the tip and receptor were monitored by continuously indenting and retracting the tip at constant lateral positions. Ｆｕｚｚｙ　ｌｏｇｉｃ  Flow chart of detecting events algorithm Convolution The integral of two functions f and g. The resulting amount overlap that occurs when g is shifted over f is the convolution of f and g. f * g ≡ ∫ f (τ ) g (t ? τ )dτ Convolution between force curves and the filter (difference) Convolution with different filters Ｃｈａｒａｃｔｅｒｉｓｔｉｃ　Ｒｅｔｒａｃｔ　ｃｕｒｖｅ Unbinding force distribution histogramsＰｅａｋ＝123.1ｐＮ，　Ａｖｅｒａｇｅ＝195.3ｐＮ Determining elastic modulus Elastic modulus is the measure of a substances tendency to be deformed elastically when a force is applied to it. E=stress/strain Elastic Modulus wave ?? ? ? ? ? ?E 0 241.46 7 298.3 15 259.37 23? ? 313.26 31 332.96 39 276.49 47 158.73 55 275.02 63 294.34? ? ? ? Average 267.0838 Results Poisson statistical method: The number of receptor-antibody pairs (n) is finite and follows a Poisson distribution. μn= σn2 F= measured force for n pairs, F = nFs , Fs= single pair force μF= μnFs, σF2 = σn2 Fs2, Fs = σF2 / μF If there are nonspecific interactions (Fn), μF= μnFs + Fn, σF2 = σn2 Fs2 = (μF- Fn) Fs= μF Fs - Fn Fs Variance versus mean of the unbinding forces Conclusion Based on the Poisson statistical method, the force required to unbind a single ligand and its pair is 123 pN and the nonspecific unbinding force is 93 pN. The unbinding force calculated is consistent with those obtained for antigen-antibody pairs b