domain swapping and different oligomeric states for the complex between calmodulin and the calmodulin-binding domain of calcineurin a域交换和不同低聚物的状态之间的复杂的钙调蛋白和钙调磷酸酶的calmodulin-binding域.pdfVIP

Domain Swapping and Different Oligomeric States for the Complex Between Calmodulin and the Calmodulin- Binding Domain of Calcineurin A Viivi Majava, Petri Kursula* Department of Biochemistry, University of Oulu, Oulu, Finland Abstract Background: Calmodulin (CaM) is a ubiquitously expressed calcium sensor that engages in regulatory interactions with a large number of cellular proteins. Previously, a unique mode of CaM target recognition has been observed in the crystal structure of a complex between CaM and the CaM-binding domain of calcineurin A. Methodology/Principal Findings: We have solved a high-resolution crystal structure of a complex between CaM and the CaM-binding domain of calcineurin A in a novel crystal form, which shows a dimeric assembly of calmodulin, as observed before in the crystal state. We note that the conformation of CaM in this complex is very similar to that of unliganded CaM, and a detailed analysis revels that the CaM-binding motif in calcineurin A is of a novel ‘1-11’ type. However, using small- angle X-ray scattering (SAXS), we show that the complex is fully monomeric in solution, and a structure of a canonically collapsed CaM-peptide complex can easily be fitted into the SAXS data. This result is also supported by size exclusion chromatography, where the addition of the ligand peptide decreases the apparent size of CaM. In addition, we studied the energetics of binding by isothermal titration calorimetry and found them to closely resemble those observed previously for ligand peptides from CaM-dependent kinases. Conclusions/Significance: Our results implicate that CaM can also form a complex with the CaM-binding domain of calcineurin in a 1:1 stoichiometr