Immunodetection of an arrestin-like protein in human retinal pigment epithelium

Christopher A. Reising, Brian Kennedy, Rita K. Getz, Nancy J. Mangini

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The goal of this study was to determine if an arrestin/S-antigen-like protein is produced by human retinal pigment epithelial (HRPE) cells maintained in tissue culture. Arrestin immunoreactivity was examined in fixed, monolayer cultures of HRPE and on immunoblots of SDS-PAGE separations of whole cell lysates of HRPE using five monoclonal antibodies (mAbs A2G5, A9C6, 3C4.2, 3D1.2 and 5c6.47) that bind to different epitopes in bovine retinal S-antigen. Monolayers of HRPE cells showed immunoreactivity with four of the mAbs though the relative staining intensity varied among mAbs and donors. For example, mAb A2G5 which historically shows very limited crossreactivity among species, reacted strongly with cells from one donor, moderately with cells from a second donor and only weakly with other donor cultures examined. mAb 3D1.2 showed no reactivity with HRPE cells. Immunoblots of SDS-PAGE separations of whole cell lysates of HRPE established from ten different donors confirmed the presence of an arrestin-related polypeptide that comigrated with retinal arrestin. These results demonstrate the presence of an arrestin-like protein in HRPE cells which have been maintained in tissue culture. Though the function of this arrestin homologue in HRPE is not yet established, it could play a role in the downregulation of receptor and/or transport protein activity.

Original languageEnglish (US)
Pages (from-to)9-15
Number of pages7
JournalCurrent Eye Research
Volume15
Issue number1
StatePublished - Jan 1996

Fingerprint

Arrestin
Retinal Pigments
Retinal Pigment Epithelium
Proteins
Epithelial Cells
Cell Separation
Polyacrylamide Gel Electrophoresis
Epitopes
Carrier Proteins
Down-Regulation
Monoclonal Antibodies
Staining and Labeling
Antigens
Peptides

Keywords

  • Arrestin
  • Cultured cells
  • HRPE
  • Immunolocalization
  • S-antigen

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

Immunodetection of an arrestin-like protein in human retinal pigment epithelium. / Reising, Christopher A.; Kennedy, Brian; Getz, Rita K.; Mangini, Nancy J.

In: Current Eye Research, Vol. 15, No. 1, 01.1996, p. 9-15.

Research output: Contribution to journalArticle

Reising, Christopher A. ; Kennedy, Brian ; Getz, Rita K. ; Mangini, Nancy J. / Immunodetection of an arrestin-like protein in human retinal pigment epithelium. In: Current Eye Research. 1996 ; Vol. 15, No. 1. pp. 9-15.
@article{cf00a02c94cc49f9808413b7ee3fbfa2,
title = "Immunodetection of an arrestin-like protein in human retinal pigment epithelium",
abstract = "The goal of this study was to determine if an arrestin/S-antigen-like protein is produced by human retinal pigment epithelial (HRPE) cells maintained in tissue culture. Arrestin immunoreactivity was examined in fixed, monolayer cultures of HRPE and on immunoblots of SDS-PAGE separations of whole cell lysates of HRPE using five monoclonal antibodies (mAbs A2G5, A9C6, 3C4.2, 3D1.2 and 5c6.47) that bind to different epitopes in bovine retinal S-antigen. Monolayers of HRPE cells showed immunoreactivity with four of the mAbs though the relative staining intensity varied among mAbs and donors. For example, mAb A2G5 which historically shows very limited crossreactivity among species, reacted strongly with cells from one donor, moderately with cells from a second donor and only weakly with other donor cultures examined. mAb 3D1.2 showed no reactivity with HRPE cells. Immunoblots of SDS-PAGE separations of whole cell lysates of HRPE established from ten different donors confirmed the presence of an arrestin-related polypeptide that comigrated with retinal arrestin. These results demonstrate the presence of an arrestin-like protein in HRPE cells which have been maintained in tissue culture. Though the function of this arrestin homologue in HRPE is not yet established, it could play a role in the downregulation of receptor and/or transport protein activity.",
keywords = "Arrestin, Cultured cells, HRPE, Immunolocalization, S-antigen",
author = "Reising, {Christopher A.} and Brian Kennedy and Getz, {Rita K.} and Mangini, {Nancy J.}",
year = "1996",
month = "1",
language = "English (US)",
volume = "15",
pages = "9--15",
journal = "Current Eye Research",
issn = "0271-3683",
publisher = "Informa Healthcare",
number = "1",

}

TY - JOUR

T1 - Immunodetection of an arrestin-like protein in human retinal pigment epithelium

AU - Reising, Christopher A.

AU - Kennedy, Brian

AU - Getz, Rita K.

AU - Mangini, Nancy J.

PY - 1996/1

Y1 - 1996/1

N2 - The goal of this study was to determine if an arrestin/S-antigen-like protein is produced by human retinal pigment epithelial (HRPE) cells maintained in tissue culture. Arrestin immunoreactivity was examined in fixed, monolayer cultures of HRPE and on immunoblots of SDS-PAGE separations of whole cell lysates of HRPE using five monoclonal antibodies (mAbs A2G5, A9C6, 3C4.2, 3D1.2 and 5c6.47) that bind to different epitopes in bovine retinal S-antigen. Monolayers of HRPE cells showed immunoreactivity with four of the mAbs though the relative staining intensity varied among mAbs and donors. For example, mAb A2G5 which historically shows very limited crossreactivity among species, reacted strongly with cells from one donor, moderately with cells from a second donor and only weakly with other donor cultures examined. mAb 3D1.2 showed no reactivity with HRPE cells. Immunoblots of SDS-PAGE separations of whole cell lysates of HRPE established from ten different donors confirmed the presence of an arrestin-related polypeptide that comigrated with retinal arrestin. These results demonstrate the presence of an arrestin-like protein in HRPE cells which have been maintained in tissue culture. Though the function of this arrestin homologue in HRPE is not yet established, it could play a role in the downregulation of receptor and/or transport protein activity.

AB - The goal of this study was to determine if an arrestin/S-antigen-like protein is produced by human retinal pigment epithelial (HRPE) cells maintained in tissue culture. Arrestin immunoreactivity was examined in fixed, monolayer cultures of HRPE and on immunoblots of SDS-PAGE separations of whole cell lysates of HRPE using five monoclonal antibodies (mAbs A2G5, A9C6, 3C4.2, 3D1.2 and 5c6.47) that bind to different epitopes in bovine retinal S-antigen. Monolayers of HRPE cells showed immunoreactivity with four of the mAbs though the relative staining intensity varied among mAbs and donors. For example, mAb A2G5 which historically shows very limited crossreactivity among species, reacted strongly with cells from one donor, moderately with cells from a second donor and only weakly with other donor cultures examined. mAb 3D1.2 showed no reactivity with HRPE cells. Immunoblots of SDS-PAGE separations of whole cell lysates of HRPE established from ten different donors confirmed the presence of an arrestin-related polypeptide that comigrated with retinal arrestin. These results demonstrate the presence of an arrestin-like protein in HRPE cells which have been maintained in tissue culture. Though the function of this arrestin homologue in HRPE is not yet established, it could play a role in the downregulation of receptor and/or transport protein activity.

KW - Arrestin

KW - Cultured cells

KW - HRPE

KW - Immunolocalization

KW - S-antigen

UR - http://www.scopus.com/inward/record.url?scp=0030052299&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030052299&partnerID=8YFLogxK

M3 - Article

C2 - 8631209

AN - SCOPUS:0030052299

VL - 15

SP - 9

EP - 15

JO - Current Eye Research

JF - Current Eye Research

SN - 0271-3683

IS - 1

ER -