Giebing T, Obermann W M, Fürst D, D'Haese J
Institut für Zoomorphologie, Zellbiologie und Parasitologie, Heinrich-Heine-Universität, Düsseldorf, Germany.
FEBS Lett. 1997 Nov 10;417(2):191-5. doi: 10.1016/s0014-5793(97)01230-1.
C- and N-terminally truncated fragments of earthworm gelsolin were constructed, cloned and expressed in Escherichia coli. G-actin-binding properties of these fragments and their influences on the polymeric state of actin were investigated. A construct lacking a large part of the third segment [E(1-295)] supports actin nucleation similar to the complete protein and shows reduced actin fragmentation property, but is no longer Ca2+-sensitive in its activity. The first and the second segments (E1 and E2) each contain one actin-binding site. In contrast to human gelsolin, E1 in combination with a short N-terminal region of E2 is not sufficient for the F-actin-severing activity of the protein.
构建了蚯蚓凝溶胶蛋白的C端和N端截短片段，将其克隆并在大肠杆菌中表达。研究了这些片段的G-肌动蛋白结合特性及其对肌动蛋白聚合状态的影响。缺失第三段大部分区域的构建体[E(1-295)]支持与完整蛋白相似的肌动蛋白成核作用，且肌动蛋白片段化特性降低，但其活性不再对Ca2+敏感。第一段和第二段（E1和E2）各包含一个肌动蛋白结合位点。与人类凝溶胶蛋白不同，E1与E2的短N端区域组合不足以实现该蛋白的F-肌动蛋白切割活性。
