Co-reconstitution and co-crystallization of phospholamban and Ca(2+)-ATPase.

Abstract:

:Significant advances have recently been made in understanding the regulation of Ca(2+)-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 Ca(2+)-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 Ca(2+)-ATPase has recently been solved by cryoelectron microscopy at 8-A resolution, thus revealing transmembrane alpha-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 Ca(2+)-ATPase at a similar level of detail.

journal_name

Ann N Y Acad Sci

authors

Young HS,Reddy LG,Jones LR,Stokes DL

doi

10.1111/j.1749-6632.1998.tb08260.x

subject

Has Abstract

pub_date

1998-09-16 00:00:00

pages

103-15

eissn

0077-8923

issn

1749-6632

journal_volume

853

pub_type

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