Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.

Rods and cones are both photoreceptors in the retina, but they are different in many aspects including the light response characteristics and, for example, cell morphology and metabolism. These differences would be caused by differences in proteins expressed in rods and cones. To understand the mole...

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Main Authors: Takashi Fukagawa, Kazuaki Takafuji, Shuji Tachibanaki, Satoru Kawamura
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2017-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC5349680?pdf=render
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spelling doaj-1691c6126e434d5f991af29d33c44db32020-11-24T21:35:37ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01123e017390810.1371/journal.pone.0173908Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.Takashi FukagawaKazuaki TakafujiShuji TachibanakiSatoru KawamuraRods and cones are both photoreceptors in the retina, but they are different in many aspects including the light response characteristics and, for example, cell morphology and metabolism. These differences would be caused by differences in proteins expressed in rods and cones. To understand the molecular bases of these differences between rods and cones, one of the ways is to compare proteins expressed in rods and cones, and to find those expressed specifically or dominantly. In the present study, we are interested in proteins in the outer segment (OS), the site responsible for generation of rod- or cone-characteristic light responses and also the site showing different morphology between rods and cones. For this, we established a method to purify the OS and the inner segment (IS) of rods and also of cones from purified carp rods and cones, respectively, using sucrose density gradient. In particular, we were interested in proteins tightly bound to the membranes of cone OS. To identify these proteins, we analyzed proteins in some selected regions of an SDS-gel of washed membranes of the OS and the IS obtained from both rods and cones, with Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS) using a protein database constructed from carp retina. By comparing the lists of the proteins found in the OS and the IS of both rods and cones, we found some proteins present in cone OS membranes specifically or dominantly, in addition to the proteins already known to be present specifically in cone OS.http://europepmc.org/articles/PMC5349680?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Takashi Fukagawa
Kazuaki Takafuji
Shuji Tachibanaki
Satoru Kawamura
spellingShingle Takashi Fukagawa
Kazuaki Takafuji
Shuji Tachibanaki
Satoru Kawamura
Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.
PLoS ONE
author_facet Takashi Fukagawa
Kazuaki Takafuji
Shuji Tachibanaki
Satoru Kawamura
author_sort Takashi Fukagawa
title Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.
title_short Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.
title_full Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.
title_fullStr Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.
title_full_unstemmed Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.
title_sort purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2017-01-01
description Rods and cones are both photoreceptors in the retina, but they are different in many aspects including the light response characteristics and, for example, cell morphology and metabolism. These differences would be caused by differences in proteins expressed in rods and cones. To understand the molecular bases of these differences between rods and cones, one of the ways is to compare proteins expressed in rods and cones, and to find those expressed specifically or dominantly. In the present study, we are interested in proteins in the outer segment (OS), the site responsible for generation of rod- or cone-characteristic light responses and also the site showing different morphology between rods and cones. For this, we established a method to purify the OS and the inner segment (IS) of rods and also of cones from purified carp rods and cones, respectively, using sucrose density gradient. In particular, we were interested in proteins tightly bound to the membranes of cone OS. To identify these proteins, we analyzed proteins in some selected regions of an SDS-gel of washed membranes of the OS and the IS obtained from both rods and cones, with Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS) using a protein database constructed from carp retina. By comparing the lists of the proteins found in the OS and the IS of both rods and cones, we found some proteins present in cone OS membranes specifically or dominantly, in addition to the proteins already known to be present specifically in cone OS.
url http://europepmc.org/articles/PMC5349680?pdf=render
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