中山大学《细胞生物学》思考题 09.docVIP

Testing Thinking of Cell Biology 2009.09.28（缺2、3） 1、You are using a cameleon indicator to measure intracellular concentrations of Ca2+．The indicator is composed of a central calmodulin domain, which converts from an extended form to a much more compact form upon calcium binding，and two flanking fluorescent proteins，with CFP attached at one end and YFP attached at the other．You have expressed this indicator in cells and now wish to measure intracellular changes in Ca2+ concentration in response to the biological process you are studying． The instructions say to excite the cameleon at 440 nm and to measure emission at 535 nm．How do you suppose the cameleon indicator works? ◆知识点： ① 报告蛋白 ② CFP（Blue,Cyan)、GFP和YFP ③ 钙调蛋白 ④ 发射光谱（emission spectra） ⑤ 激发光谱（excitation spectra） ◆解题思维（分析能力）： ① CFP与YFP的作用 ② Ca2+与钙调蛋白的关系 ◆本题的来源：Miyawaki A, Llopis J, Heim R, McCafferty JM, Adams JA, Ikura M Tsien RY (1997) Fluorescent indicators for Ca2+ based on green fluorescent protein and calmodulin. Nature 388, 882–887. 2009.10.12 1、You are studying the binding of proteins to the cytoplasmic face of cultured neuroblastoma (神经母细胞瘤) cells and have found a method that gives a good yield of inside-out vesicles from the plasma membrane. Unfortunately, your preparations are contaminated with variable amounts of right-side-out vesicles. Nothing you have tried avoids this variable contamination. A friend suggests that you pass your vesicles over an affinity column made of lectin coupled to solid beads. What is the point of your friend’s suggestion? 回答关键词：膜糖仅存在于外膜，凝集素与糖亲和，亲和层析的方法。 2、Gorter and Grendel Revisited. Gorter and Grendels classic conclusion that the plasma membrane of the human erythrocyte consists of a lipid bilayer was based on the following observations:(i) the lipids that they extracted with acetone from 4.74 ×109 erythrocytes formed a monolayer 0.89 m2 in area when spread out on a water surface; and (ii) the surface area of one erythrocyte was about 100μm2, according