Webbläsaren som du använder stöds inte av denna webbplats. Alla versioner av Internet Explorer stöds inte längre, av oss eller Microsoft (läs mer här: * https://www.microsoft.com/en-us/microsoft-365/windows/end-of-ie-support).

Var god och använd en modern webbläsare för att ta del av denna webbplats, som t.ex. nyaste versioner av Edge, Chrome, Firefox eller Safari osv.

Double Electron-Electron Resonance Probes Ca2+-Induced Conformational Changes and Dimerization of Recoverin

In English

Författare

  • William K. Myers
  • Xianzhong Xu
  • Congmin Li
  • Jens Lagerstedt
  • Madhu S. Budamagunta
  • John C. Voss
  • R. David Britt
  • James B. Ames

Summary, in English

Recoverin a member of the neuronal calcium sensor (NCS) branch of the calmodulin superfamily, is expressed in retinal photoreceptor cells and serves as a calcium sensor in vision. Ca2+-induced conformational changes in recoverin cause extrusion of its covalently attached myristate (termed Ca2+-myristoyl switch) that promotes translocation of recoverin to disk membranes during phototransduction in retinal rod cells. Here we report double electron electron resonance (DEER) experiments on recoverin that probe Ca2+-induced changes in distance as measured by the dipolar coupling between spin-labels strategically positioned at engineered cysteine residues on the protein surface. The DEER distance between nitroxide spin-labels attached at C39 and N120C is 2.5 +/- 0.1 nm for Ca2+-free recoverin and 3.7 +/- 0.1 nm for Ca2+-bound recoverin. An additional DEER distance (5-6 nm) observed for Ca2+-bound recoverin may represent an intermolecular distance between C39 and N120. N-15 NMR relaxation analysis and CW-EPR experiments both confirm that Ca2+-bound recoverin forms a dimer at protein concentrations above 100 mu M, whereas Ca2+-free recoverin is monomeric We propose that Ca2+-induced dimerization of recoverin at the disk membrane surface may play a role in regulating Ca2+-dependent phosphorylation of dimeric rhodopsin. The DEER approach will be useful for elucidating dimeric structures of NCS proteins in general for which Ca2+-induced dimerization is functionally important but not well understood.

Avdelning/ar

Publiceringsår

2013

Språk

Engelska

Sidor

5800-5808

Publikation/Tidskrift/Serie

Biochemistry

Volym

52

Issue

34

Dokumenttyp

Artikel i tidskrift

Förlag

The American Chemical Society (ACS)

Ämne

  • Biochemistry and Molecular Biology

Status

Published

Forskningsgrupp

  • Medical Protein Science

ISBN/ISSN/Övrigt

  • ISSN: 0006-2960