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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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Autores principales: Fukagawa, Takashi, Takafuji, Kazuaki, Tachibanaki, Shuji, Kawamura, Satoru
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5349680/
https://www.ncbi.nlm.nih.gov/pubmed/28291804
http://dx.doi.org/10.1371/journal.pone.0173908
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author Fukagawa, Takashi
Takafuji, Kazuaki
Tachibanaki, Shuji
Kawamura, Satoru
author_facet Fukagawa, Takashi
Takafuji, Kazuaki
Tachibanaki, Shuji
Kawamura, Satoru
author_sort Fukagawa, Takashi
collection PubMed
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.
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spelling pubmed-53496802017-04-06 Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina Fukagawa, Takashi Takafuji, Kazuaki Tachibanaki, Shuji Kawamura, Satoru PLoS One Research Article 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. Public Library of Science 2017-03-14 /pmc/articles/PMC5349680/ /pubmed/28291804 http://dx.doi.org/10.1371/journal.pone.0173908 Text en © 2017 Fukagawa et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Fukagawa, Takashi
Takafuji, Kazuaki
Tachibanaki, Shuji
Kawamura, Satoru
Purification of cone outer segment for proteomic analysis on its membrane proteins in carp retina
title 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_short 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
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5349680/
https://www.ncbi.nlm.nih.gov/pubmed/28291804
http://dx.doi.org/10.1371/journal.pone.0173908
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