Identification of the retinal pigment epithelium protein RET-PE2 as CE- 9/OX-47, a member of the immunoglobulin superfamily

Silvia C. Finnemann, Alan D Marmorstein, James M. Neill, Enrique Rodriguez-Boulan

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Purpose. To identify the retinal pigment epithelium (RPE) surface antigen recognized by the monoclonal antibody RET-PE2. Methods. A lambda bacteriophage complementary DNA (cDNA) expression library, representing the rat RPE cell line RPE-J, was constructed and screened with the RET-PE2 monoclonal antibody. Transient transfections of the RET-PE2 cDNA, immunofluorescence stainings of tissue sections or cultured cells, and Western blot analyses of tissue and cell detergent extracts served to prove that the protein resulting from expression of the cDNA is the RET-PE2 antigen. Results. Three independent cDNAs were cloned that shared overlapping sequences. Sequence alignment with EMBL database entries revealed identity to the published cDNA of CE-9/OX-47, a member of the immunoglobulin superfamily. One of the clones encoded the entire open reading frame of CE-9. The expression pattern of the RET-PE2 antigen matched that of CE-9, which is widely expressed. Chinese hamster ovary cells transiently transfected with the RET-PE2 cDNA produced a membrane-localized protein that was recognized by RET-PE2 and CE-9 antibodies. Conclusions. The antibody RET-PE2 recognizes the CE-9/OX-47 gene product, a transmembrane protein of the immunoglobulin superfamily. Contrary to results reported earlier, RET-PE2 immunoreactivity is widely distributed among different rat tissues-kidney, liver, and testis. In epithelia other than the adult RPE, it is confined to the basolateral plasma membrane. Its apical polarization in the RPE of adult rats supports earlier findings that some proteins that are basolateral in other epithelia exhibit reversed polarity in the RPE.

Original languageEnglish (US)
Pages (from-to)2366-2374
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number11
StatePublished - Oct 1997
Externally publishedYes

Fingerprint

Retinal Pigment Epithelium
Immunoglobulins
Complementary DNA
Proteins
Epithelium
Monoclonal Antibodies
Bacteriophage lambda
Antigens
Antibodies
Sequence Alignment
Surface Antigens
Cricetulus
Cell Extracts
Gene Library
Detergents
Open Reading Frames
Fluorescent Antibody Technique
Transfection
Testis
Ovary

Keywords

  • Antigen
  • Apical surface proteins
  • Cell polarity
  • Gene expression
  • RET-PE2
  • Retinal pigment epithelium

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Identification of the retinal pigment epithelium protein RET-PE2 as CE- 9/OX-47, a member of the immunoglobulin superfamily. / Finnemann, Silvia C.; Marmorstein, Alan D; Neill, James M.; Rodriguez-Boulan, Enrique.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 11, 10.1997, p. 2366-2374.

Research output: Contribution to journalArticle

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abstract = "Purpose. To identify the retinal pigment epithelium (RPE) surface antigen recognized by the monoclonal antibody RET-PE2. Methods. A lambda bacteriophage complementary DNA (cDNA) expression library, representing the rat RPE cell line RPE-J, was constructed and screened with the RET-PE2 monoclonal antibody. Transient transfections of the RET-PE2 cDNA, immunofluorescence stainings of tissue sections or cultured cells, and Western blot analyses of tissue and cell detergent extracts served to prove that the protein resulting from expression of the cDNA is the RET-PE2 antigen. Results. Three independent cDNAs were cloned that shared overlapping sequences. Sequence alignment with EMBL database entries revealed identity to the published cDNA of CE-9/OX-47, a member of the immunoglobulin superfamily. One of the clones encoded the entire open reading frame of CE-9. The expression pattern of the RET-PE2 antigen matched that of CE-9, which is widely expressed. Chinese hamster ovary cells transiently transfected with the RET-PE2 cDNA produced a membrane-localized protein that was recognized by RET-PE2 and CE-9 antibodies. Conclusions. The antibody RET-PE2 recognizes the CE-9/OX-47 gene product, a transmembrane protein of the immunoglobulin superfamily. Contrary to results reported earlier, RET-PE2 immunoreactivity is widely distributed among different rat tissues-kidney, liver, and testis. In epithelia other than the adult RPE, it is confined to the basolateral plasma membrane. Its apical polarization in the RPE of adult rats supports earlier findings that some proteins that are basolateral in other epithelia exhibit reversed polarity in the RPE.",
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T1 - Identification of the retinal pigment epithelium protein RET-PE2 as CE- 9/OX-47, a member of the immunoglobulin superfamily

AU - Finnemann, Silvia C.

AU - Marmorstein, Alan D

AU - Neill, James M.

AU - Rodriguez-Boulan, Enrique

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N2 - Purpose. To identify the retinal pigment epithelium (RPE) surface antigen recognized by the monoclonal antibody RET-PE2. Methods. A lambda bacteriophage complementary DNA (cDNA) expression library, representing the rat RPE cell line RPE-J, was constructed and screened with the RET-PE2 monoclonal antibody. Transient transfections of the RET-PE2 cDNA, immunofluorescence stainings of tissue sections or cultured cells, and Western blot analyses of tissue and cell detergent extracts served to prove that the protein resulting from expression of the cDNA is the RET-PE2 antigen. Results. Three independent cDNAs were cloned that shared overlapping sequences. Sequence alignment with EMBL database entries revealed identity to the published cDNA of CE-9/OX-47, a member of the immunoglobulin superfamily. One of the clones encoded the entire open reading frame of CE-9. The expression pattern of the RET-PE2 antigen matched that of CE-9, which is widely expressed. Chinese hamster ovary cells transiently transfected with the RET-PE2 cDNA produced a membrane-localized protein that was recognized by RET-PE2 and CE-9 antibodies. Conclusions. The antibody RET-PE2 recognizes the CE-9/OX-47 gene product, a transmembrane protein of the immunoglobulin superfamily. Contrary to results reported earlier, RET-PE2 immunoreactivity is widely distributed among different rat tissues-kidney, liver, and testis. In epithelia other than the adult RPE, it is confined to the basolateral plasma membrane. Its apical polarization in the RPE of adult rats supports earlier findings that some proteins that are basolateral in other epithelia exhibit reversed polarity in the RPE.

AB - Purpose. To identify the retinal pigment epithelium (RPE) surface antigen recognized by the monoclonal antibody RET-PE2. Methods. A lambda bacteriophage complementary DNA (cDNA) expression library, representing the rat RPE cell line RPE-J, was constructed and screened with the RET-PE2 monoclonal antibody. Transient transfections of the RET-PE2 cDNA, immunofluorescence stainings of tissue sections or cultured cells, and Western blot analyses of tissue and cell detergent extracts served to prove that the protein resulting from expression of the cDNA is the RET-PE2 antigen. Results. Three independent cDNAs were cloned that shared overlapping sequences. Sequence alignment with EMBL database entries revealed identity to the published cDNA of CE-9/OX-47, a member of the immunoglobulin superfamily. One of the clones encoded the entire open reading frame of CE-9. The expression pattern of the RET-PE2 antigen matched that of CE-9, which is widely expressed. Chinese hamster ovary cells transiently transfected with the RET-PE2 cDNA produced a membrane-localized protein that was recognized by RET-PE2 and CE-9 antibodies. Conclusions. The antibody RET-PE2 recognizes the CE-9/OX-47 gene product, a transmembrane protein of the immunoglobulin superfamily. Contrary to results reported earlier, RET-PE2 immunoreactivity is widely distributed among different rat tissues-kidney, liver, and testis. In epithelia other than the adult RPE, it is confined to the basolateral plasma membrane. Its apical polarization in the RPE of adult rats supports earlier findings that some proteins that are basolateral in other epithelia exhibit reversed polarity in the RPE.

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