Drosophila Neurocalcin, a Fatty Acylated, Ca-binding Protein that Associates with Membranes and Inhibits in Vitro Phosphorylation of Bovine Rhodopsin (*)
Drosophila Neurocalcin, a Fatty Acylated, Ca-binding Protein that Associates with Membranes and Inhibits in Vitro Phosphorylation of Bovine Rhodopsin (*)
复制标题
果蝇神经钙蛋白，一种脂肪酰化 Ca 结合蛋白，与膜结合并抑制牛视紫红质的体外磷酸化 (*)
DOI：
--
复制
发表时间：
1996
影响因子：
4.8
通讯作者：
DAVID H. F. Teng
中科院分区：
文献类型：
--
作者：
E. Faurobert;Ching;J. Hurley;DAVID H. F. Teng
神经钙蛋白是一类神经元特异性的钙结合蛋白家族。在此之前，我们报道了果蝇神经钙蛋白的克隆和初步鉴定（Teng，D. H.- F.、陈春K.，和Hurley，J. B.（1994）J.Biol.Chem.269，31900-31907）。我们发现，果蝇神经钙蛋白蛋白（DrosNCa）在神经元中表达，并且细菌表达的重组DrosNCa（rDrosNCa）可以被肉豆蔻酰化。在这里，我们提出了两条证据表明DrosNCa在体内是脂肪酰化的。首先，亲和纯化的天然DrosNCa在二维凝胶电泳上的迁移率与肉豆蔻酰化rDrosNCa相同，而与非酰化rDrosNCa不同。其次，天然DrosNCa的膜结合特性与肉豆蔻酰化rDrosNCa的膜结合特性相似;这两种蛋白质在0.2 mM Ca时均与膜结合，而非酰化rDrosNCa始终保持可溶性。已经表明，当Ca存在时，恢复蛋白抑制视紫红质的磷酸化（Kawamura等人，1993），并且依赖性恢复蛋白/视紫红质激酶相互作用是恢复蛋白抑制作用的基础（Chen et al.，1995年）。鉴于恢复蛋白和神经钙蛋白之间的相似性，我们研究了DrosNCa对视紫红质磷酸化的影响。我们发现，rDrosNCa是能够抑制牛视紫红质磷酸化在体外钙依赖的方式。豆蔻酰化增强rDrosNCa的抑制活性，其作用的效力与恢复蛋白的效力相似。其他两个相关的EF手蛋白，鸟苷酸环化酶激活蛋白-2和钙调蛋白，在这些磷酸化测定中仅是差的抑制剂。因此，视紫红质磷酸化的体外抑制似乎是恢复素样蛋白亚类的可测定性质。
Neurocalcins belong to a family of neuronal specific EF hand Ca-binding proteins defined by recoverin. Previously, we reported the cloning and initial characterization of neurocalcin in Drosophila melanogaster (Teng, D. H.-F., Chen, C.-K., and Hurley, J. B.(1994) J. Biol. Chem. 269, 31900-31907). We showed that the Drosophila neurocalcin protein (DrosNCa) is expressed in neurons and that bacterially expressed recombinant DrosNCa (rDrosNCa) can be myristoylated. Here, we present two lines of evidence that DrosNCa is fatty acylated in vivo. First, the mobility of affinity-purified native DrosNCa on two-dimensional gel electrophoresis is identical to that of myristoylated rDrosNCa and distinct from that of nonacylated rDrosNCa. Second, the membrane binding properties of native DrosNCa are similar to those of myristoylated rDrosNCa; both of these proteins bind to membranes at 0.2 mM Ca, whereas nonacylated rDrosNCa always remains soluble. It has been shown that recoverin inhibits the phosphorylation of rhodopsin when Ca is present (Kawamura et al., 1993) and that a dependent recoverin/rhodopsin kinase interaction underlies the inhibitory effect of recoverin (Chen et al., 1995). Given the similarities between recoverin and neurocalcin, we examined the effect of DrosNCa on rhodopsin phosphorylation. We find that rDrosNCa is capable of inhibiting bovine rhodopsin phosphorylation in vitro in a Ca-dependent manner. The inhibitory activity of rDrosNCa is enhanced by myristoylation, and the potency of its effect is similar to that of recoverin. Two other related EF hand proteins, guanylate cyclase-activating protein-2 and calmodulin, are only poor inhibitors in these phosphorylation assays. in vitro inhibition of rhodopsin phosphorylation therefore appears to be an assayable property of a subset of recoverin-like proteins.
视网膜恢复素的 NH2 末端被一小类脂肪酸酰化。
DOI：
--
发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Dizhoor,AM;Ericsson,LH;Johnson,RS;Kumar,S;Olshevskaya,E;Zozulya,S;Neubert,TA;Stryer,L;Hurley,JB;Walsh,KA
通讯作者：
Walsh,KA
视觉钙传感器恢复蛋白的克隆、表达和结晶。
DOI：
10.1073/pnas.89.13.5705
发表时间：
1992
影响因子：
11.1
作者：
Ray,S;Zozulya,S;Niemi,GA;Flaherty,KM;Brolley,D;Dizhoor,AM;McKay,DB;Hurley,J;Stryer,L
通讯作者：
Stryer,L
恢复素肉豆蔻酰基的钙依赖性溶剂化。
影响因子：
2.9
作者：
Hughes,RE;Brzovic,PS;Klevit,RE;Hurley,JB
通讯作者：
Hurley,JB