> Detailanzeige

Hou, Sing‐Yi; Govorunova, Elena G.; Ntefidou, Maria; Lane, C. Elizabeth; Spudich, Elena N.; Sineshchekov, Oleg A.; Spudich, John L.

Diversity of Chlamydomonas Channelrhodopsins

Literatur-
verwaltung

Zur
Merkliste

Lösche von
Merkliste

Per Whatsapp teilen

Schließen

Merkliste



Sie können Bookmarks mittels Listen verwalten, loggen Sie sich dafür bitte in Ihr SLUB Benutzerkonto ein.
  • Medientyp: E-Artikel
  • Titel: Diversity of Chlamydomonas Channelrhodopsins
  • Beteiligte: Hou, Sing‐Yi; Govorunova, Elena G.; Ntefidou, Maria; Lane, C. Elizabeth; Spudich, Elena N.; Sineshchekov, Oleg A.; Spudich, John L.
  • Erschienen: Wiley, 2012
  • Erschienen in: Photochemistry and Photobiology
  • Sprache: Englisch
  • DOI: 10.1111/j.1751-1097.2011.01027.x
  • ISSN: 0031-8655; 1751-1097
  • Entstehung:
  • Anmerkungen:
  • Beschreibung: <jats:title>Abstract</jats:title><jats:p>Channelrhodopsins act as photoreceptors for control of motility behavior in flagellates and are widely used as genetically targeted tools to optically manipulate the membrane potential of specific cell populations (“optogenetics”). The first two channelrhodopsins were obtained from the model organism <jats:italic>Chlamydomonas reinhardtii</jats:italic> (<jats:italic>Cr</jats:italic>ChR1 and <jats:italic>Cr</jats:italic>ChR2). By homology cloning we identified three new channelrhodopsin sequences from the same genus, <jats:italic>Ca</jats:italic>ChR1, <jats:italic>Cy</jats:italic>ChR1 and <jats:italic>Cra</jats:italic>ChR2, from <jats:italic>C. augustae, C. yellowstonensis</jats:italic> and <jats:italic>C. raudensis</jats:italic>, respectively. <jats:italic>Ca</jats:italic>ChR1 and <jats:italic>Cy</jats:italic>ChR1 were functionally expressed in HEK293 cells, where they acted as light‐gated ion channels similar to <jats:italic>Cr</jats:italic>ChR1. However, both, which are similar to each other, differed from <jats:italic>Cr</jats:italic>ChR1 in current kinetics, inactivation, light intensity dependence, spectral sensitivity and dependence on the external pH. These results show that extensive channelrhodopsin diversity exists even within the same genus, <jats:italic>Chlamydomonas</jats:italic>. The maximal spectral sensitivity of <jats:italic>Ca</jats:italic>ChR1 was at 520 nm at pH 7.4, about 40 nm redshifted as compared to that of <jats:italic>Cr</jats:italic>ChR1 under the same conditions. <jats:italic>Ca</jats:italic>ChR1 was successfully expressed in <jats:italic>Pichia pastoris</jats:italic> and exhibited an absorption spectrum identical to the action spectrum of <jats:italic>Ca</jats:italic>ChR1‐generated photocurrents. The redshifted spectra and the lack of fast inactivation in <jats:italic>Ca</jats:italic>ChR1‐ and <jats:italic>Cy</jats:italic>ChR1‐generated currents are features desirable for optogenetics applications.</jats:p>