A second rhodopsin-like protein in Cyanophora paradoxa: Gene sequence and protein expression in a cell-free system
Authors: Frassanito A. M., Barsanti L., Passarelli V., Evangelista V., Gualtieri P.
Journal: Journal of Photochemistry and Photobiology B-Biology
DOI: 10.1016/j.jphotobiol.2013.06.010
Abstract:
Here we report the identification and expression of a second rhodopsin-like protein in the alga Cyanophora paradoxa (Glaucophyta), named Cyanophopsin_2. This new protein was identified due to a serendipity event, since the RACE reaction performed to complete the sequence of Cyanophopsin_1, (the first rhodopsin-like protein of C. paradoxa identified in 2009 by our group), amplified a 619 bp sequence corresponding to a portion of a new gene of the same protein family. The full sequence consists of 1175 bp consisting of 849 bp coding DNA sequence and 4 introns of 326 bp. The protein is characterized by an N-terminal region of 47 amino acids, followed by a region with 7 α-helices of 213 amino acids and a C-terminal region of 22 amino acids. This protein showed high identity with Cyanophopsin_1 and other rhodopsin-like proteins of Archea, Bacteria, Fungi and Algae. Cyanophosin_2 (CpR2) was expressed in a cell-free expression system, and characterized by means of absorption spectroscopy.
Keywords: Rhodopsin-like protein, Cyanophora paradoxa, CpR2, Cell-free expression, Absorption spectrum
