Co-reconstitution and co-crystallization of phospholamban and Ca(2+)-ATPase.
Co-reconstitution and co-crystallization of phospholamban and Ca(2+)-ATPase.
复制标题
受磷蛋白和 Ca(2 )-ATPase 的共重构和共结晶。
DOI：
10.1111/j.1749-6632.1998.tb08260.x
复制
发表时间：
1998
影响因子：
5.2
通讯作者：
Stokes,DL
中科院分区：
文献类型：
--
作者：
Young,HS;Reddy,LG;Jones,LR;Stokes,DL
Significant advances have recently been made in understanding the regulation of Ca2+‐ATPase by phospholamban and in modeling their structures. However, these insights would be furthered by determining the 3‐D structure of both proteins within the membrane, thus revealing the structural basis for their interaction. To this end, we have developed methods for reconstituting purified Ca2+‐ATPase with recombinant phospholamban. After reconstitution at high lipid‐to‐protein ratios, we have verified their functional association by measuring calcium transport and ATPase activity. Furthermore, we have grown co‐crystals after reconstitution at low lipid‐to‐protein ratios. The structure of Ca2+‐ATPase has recently been solved by cryoelectron microscopy at 8‐Å resolution, thus revealing transmembrane α‐helices. Using a variety of constraints, we have associated these helices with the predicted transmembrane sequences to produce a detailed model for the packing of transmembrane helices. Structure determination of the co‐crystals is currently underway, which we hope will eventually reveal the interaction of phospholamban with Ca2+‐ATPase at a similar level of detail.
