% IMPORTANT: The following is UTF-8 encoded.  This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.

@ARTICLE{Gensch:56498,
      author       = {Gensch, T. and Komolov, K. E. and Senin, I. I. and
                      Philippov, P. P. and Koch, K.-W.},
      title        = {{C}a2+-dependent conformational changes in the neuronal
                      {C}a2+-sensor recoverin probed by the fluorescent dye
                      {A}lexa647},
      journal      = {Proteins},
      volume       = {66},
      issn         = {0887-3585},
      address      = {New York, NY},
      publisher    = {Wiley-Liss},
      reportid     = {PreJuSER-56498},
      pages        = {492 - 499},
      year         = {2007},
      note         = {Record converted from VDB: 12.11.2012},
      abstract     = {Recoverin belongs to the superfamily of EF-hand
                      Ca2+-binding proteins and operates as a Ca2+-sensor in
                      vertebrate photoreceptor cells, where it regulates the
                      activity of rhodopsin kinase GRK1 in a Ca2+-dependent
                      manner. Ca2+-dependent conformational changes in recoverin
                      are allosterically controlled by the covalently attached
                      myristoyl group. The amino acid sequence of recoverin
                      harbors a unique cysteine at position 38. The cysteine can
                      be modified by the fluorescent dye Alexa647 using a
                      maleimide-thiol coupling step. Introduction of Alexa647 into
                      recoverin did not disturb the biological function of
                      recoverin, as it can regulate rhodopsin kinase activity like
                      unlabeled recoverin. Performance of the Ca2+-myristoyl
                      switch of labeled recoverin was monitored by Ca2+-dependent
                      association with immobilized lipids using surface plasmon
                      resonance spectroscopy. When the Ca2+-concentration was
                      varied, labeled myristoylated recoverin showed a
                      $37\%-change$ in fluorescence emission and a $34\%-change$
                      in excitation intensity, emission and excitation maxima
                      shifted by 6 and 18 nm, respectively. In contrast, labeled
                      nonmyristoylated recoverin exhibited only minimal changes.
                      Time-resolved fluorescence measurements showed
                      biexponentiell fluorescence decay, in which the slower time
                      constant of 2 ns was specifically influenced by Ca2+-induced
                      conformational changes. A similar influence on the slower
                      time constant was observed with the recoverin mutant RecE85Q
                      that has a disabled EF-hand 2, but no such influence was
                      detected with the mutant RecE121Q (EF-hand 3 is
                      nonfunctional) that contains the myristoyl group in a
                      clamped position. We conclude from our results that Alexa647
                      bound to cysteine 38 can monitor the conformational
                      transition in recoverin that is under control of the
                      myristoyl group.},
      keywords     = {Amino Acid Substitution / Animals / Calcium: pharmacology /
                      Calcium: physiology / Cattle / Cyclic AMP: analogs $\&$
                      derivatives / Cyclic AMP: chemistry / Cysteine: chemistry /
                      Fluorescent Dyes: chemistry / G-Protein-Coupled Receptor
                      Kinase 1: metabolism / Models, Molecular / Mutagenesis,
                      Site-Directed / Mutation, Missense / Myristic Acid:
                      chemistry / Point Mutation / Protein Binding / Protein
                      Conformation / Protein Processing, Post-Translational /
                      Protein Structure, Tertiary / Recombinant Fusion Proteins:
                      chemistry / Recoverin: chemistry / Recoverin: drug effects /
                      Recoverin: genetics / Spectrometry, Fluorescence /
                      Structure-Activity Relationship / Surface Plasmon Resonance
                      / Fluorescent Dyes (NLM Chemicals) / RCV1 protein, Bos
                      taurus (NLM Chemicals) / Recombinant Fusion Proteins (NLM
                      Chemicals) / Recoverin (NLM Chemicals) / 8-aminohexylamino
                      cAMP (NLM Chemicals) / Cysteine (NLM Chemicals) / Myristic
                      Acid (NLM Chemicals) / Cyclic AMP (NLM Chemicals) / Calcium
                      (NLM Chemicals) / G-Protein-Coupled Receptor Kinase 1 (NLM
                      Chemicals) / J (WoSType)},
      cin          = {INB-1},
      ddc          = {540},
      cid          = {I:(DE-Juel1)VDB804},
      pnm          = {Funktion und Dysfunktion des Nervensystems},
      pid          = {G:(DE-Juel1)FUEK409},
      shelfmark    = {Biochemistry $\&$ Molecular Biology / Biophysics},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:17078090},
      UT           = {WOS:000243358000018},
      doi          = {10.1002/prot.21231},
      url          = {https://juser.fz-juelich.de/record/56498},
}