Calcium binding decreases the stokes radius of calmodulin and mutants R74A, R90A, and R90G.

Abstract:

:Calmodulin (CaM) is an intracellular cooperative calcium-binding protein essential for activating many diverse target proteins. Biophysical studies of the calcium-induced conformational changes of CaM disagree on the structure of the linker between domains and possible orientations of the domains. Molecular dynamics studies have predicted that Ca4(2+)CaM is in equilibrium between an extended and compact conformation and that Arg74 and Arg90 are critical to the compaction process. In this study gel permeation chromatography was used to resolve calcium-induced changes in the hydrated shape of CaM at pH 7.4 and 5.6. Results showed that mutation of Arg 74 to Ala increases the R(s) as predicted; however, the average separation of domains in Ca4(2+)-CaM was larger than predicted by molecular dynamics. Mutation of Arg90 to Ala or Gly affected the dimensions of apo-CaM more than those of Ca4(2+)-CaM. Calcium binding to CaM and mutants (R74A-CaM, R90A-CaM, and R90G-CaM) lowered the Stokes radius (R(s)). Differences between R(s) values reported here and Rg values determined by small-angle x-ray scattering studies illustrate the importance of using multiple techniques to explore the solution properties of a flexible protein such as CaM.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Sorensen BR,Shea MA

doi

10.1016/S0006-3495(96)79535-8

subject

Has Abstract

pub_date

1996-12-01 00:00:00

pages

3407-20

issue

6

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(96)79535-8

journal_volume

71

pub_type

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