Domain structure of a mammalian myosin I beta.
Domain structure of a mammalian myosin I beta.
复制标题
哺乳动物肌球蛋白 Iβ 的结构域结构。
DOI：
10.1073/pnas.91.14.6349
复制
发表时间：
1994
影响因子：
11.1
通讯作者：
Albanesi,JP
中科院分区：
文献类型：
--
作者：
Reizes,O;Barylko,B;Li,C;Südhof,TC;Albanesi,JP
我们确定了牛脑中肌球蛋白 I（称为哺乳动物肌球蛋白 I beta，MMI beta）的一级结构，并确定了其功能域。该蛋白质之前是从大脑和肾上腺中纯化的。在大肠杆菌中生成并表达了几种构建体作为谷胱甘肽 S-转移酶融合蛋白，重组蛋白被识别天然肌球蛋白 I“头”或“尾”结构域的单克隆抗体识别。使用凝胶覆盖方法来确认钙调蛋白与 MMI beta 中共有的钙调蛋白结合序列结合。结合测定用于检测与阴离子磷脂囊泡的相互作用。我们得出结论，MMI beta 由包含 ATP 和肌动蛋白结合位点的氨基末端 80.5 kDa 结构域、随后具有三个钙调蛋白结合序列的 8.5 kDa 结构域和与阴离子磷脂和膜结合的基本 30 kDa 羧基末端尾段组成。
We have determined the primary structure of a myosin I (called mammalian myosin I beta, MMI beta) from bovine brain and identified its functional domains. The protein was previously purified from brain and adrenal gland. Several constructs were generated and expressed in Escherichia coli as glutathione S-transferase fusion proteins and the recombinant proteins were recognized by monoclonal antibodies that recognize either "head" or "tail" domains of native myosin I. A gel overlay method was used to confirm that calmodulin binds to the consensus calmodulin-binding sequence in MMI beta. Binding assays were used to detect interaction with anionic phospholipid vesicles. We conclude that MMI beta consists of an amino-terminal 80.5-kDa domain that contains the ATP- and actin-binding sites, followed by an 8.5-kDa domain with three calmodulin-binding sequences and a basic 30-kDa carboxyl-terminal tail segment that binds to anionic phospholipids and membranes.