Identification of an IgE-binding protein by molecular cloning

Fu-Tong Liu, K. Albrandt, E. Mendel, A. Kulczycki, N. K. Orida

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

The synthesis and function of IgE are dependent on IgE-binding proteins, which include cell surface IgE receptors and IgE-binding lymphokines. To further our understanding of the IgE system, we have engaged in the molecular cloning of genes for some of these proteins. In studying the in vitro translation products of mRNA from rat basophilic leukemia (RBL) cells, we have identified a M(r) 31,000 polypeptide that binds IgE and is also reactive with antibodies to proteins affinity-purified from RBL cells with IgE immunoadsorbent. For the molecular cloning, double-stranded cDNA was synthesized from sucrose gradient-fractionated RBL mRNA, inserted into plasmid pBR322, and used to transform Escherichia coli. By screening transformants with a hybridization-selection/in vitro translation procedure, we identified one clone containing cDNA that hybridized to mRNA coding for a M(r) 31,000 IgE-binding protein. The DNA sequence of this cloned cDNA (571 base pairs) was determined and the amino acid sequence corresponding to part of the protein was deduced. In RNA blot analysis, the cDNA hybridized with a mRNA of 1100 nucleotides found in RBL cells but absent in cells not expressing IgE receptors. This cloned cDNA most likely codes for the M(r) 31,000 IgE-binding protein identified in RBL cells, which appears to be related to the IgE-binding phenotype of the cells and which may have a significant role in the IgE-mediated activation of basophils and mast cells.

Original languageEnglish (US)
Pages (from-to)4100-4104
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume82
Issue number12
DOIs
StatePublished - 1985

Fingerprint

Galectin 3
Molecular Cloning
Immunoglobulin E
Leukemia
Complementary DNA
IgE Receptors
Messenger RNA
Immunosorbents
Proteins
Basophils
Lymphokines
Cell Surface Receptors
Protein Biosynthesis
Mast Cells
Base Pairing
Sucrose
Amino Acid Sequence
Plasmids
Nucleotides
Clone Cells

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Identification of an IgE-binding protein by molecular cloning. / Liu, Fu-Tong; Albrandt, K.; Mendel, E.; Kulczycki, A.; Orida, N. K.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 82, No. 12, 1985, p. 4100-4104.

Research output: Contribution to journalArticle

Liu, Fu-Tong ; Albrandt, K. ; Mendel, E. ; Kulczycki, A. ; Orida, N. K. / Identification of an IgE-binding protein by molecular cloning. In: Proceedings of the National Academy of Sciences of the United States of America. 1985 ; Vol. 82, No. 12. pp. 4100-4104.
@article{c92afd59b33d428c83ec42ab1ead75ff,
title = "Identification of an IgE-binding protein by molecular cloning",
abstract = "The synthesis and function of IgE are dependent on IgE-binding proteins, which include cell surface IgE receptors and IgE-binding lymphokines. To further our understanding of the IgE system, we have engaged in the molecular cloning of genes for some of these proteins. In studying the in vitro translation products of mRNA from rat basophilic leukemia (RBL) cells, we have identified a M(r) 31,000 polypeptide that binds IgE and is also reactive with antibodies to proteins affinity-purified from RBL cells with IgE immunoadsorbent. For the molecular cloning, double-stranded cDNA was synthesized from sucrose gradient-fractionated RBL mRNA, inserted into plasmid pBR322, and used to transform Escherichia coli. By screening transformants with a hybridization-selection/in vitro translation procedure, we identified one clone containing cDNA that hybridized to mRNA coding for a M(r) 31,000 IgE-binding protein. The DNA sequence of this cloned cDNA (571 base pairs) was determined and the amino acid sequence corresponding to part of the protein was deduced. In RNA blot analysis, the cDNA hybridized with a mRNA of 1100 nucleotides found in RBL cells but absent in cells not expressing IgE receptors. This cloned cDNA most likely codes for the M(r) 31,000 IgE-binding protein identified in RBL cells, which appears to be related to the IgE-binding phenotype of the cells and which may have a significant role in the IgE-mediated activation of basophils and mast cells.",
author = "Fu-Tong Liu and K. Albrandt and E. Mendel and A. Kulczycki and Orida, {N. K.}",
year = "1985",
doi = "10.1073/pnas.82.12.4100",
language = "English (US)",
volume = "82",
pages = "4100--4104",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "12",

}

TY - JOUR

T1 - Identification of an IgE-binding protein by molecular cloning

AU - Liu, Fu-Tong

AU - Albrandt, K.

AU - Mendel, E.

AU - Kulczycki, A.

AU - Orida, N. K.

PY - 1985

Y1 - 1985

N2 - The synthesis and function of IgE are dependent on IgE-binding proteins, which include cell surface IgE receptors and IgE-binding lymphokines. To further our understanding of the IgE system, we have engaged in the molecular cloning of genes for some of these proteins. In studying the in vitro translation products of mRNA from rat basophilic leukemia (RBL) cells, we have identified a M(r) 31,000 polypeptide that binds IgE and is also reactive with antibodies to proteins affinity-purified from RBL cells with IgE immunoadsorbent. For the molecular cloning, double-stranded cDNA was synthesized from sucrose gradient-fractionated RBL mRNA, inserted into plasmid pBR322, and used to transform Escherichia coli. By screening transformants with a hybridization-selection/in vitro translation procedure, we identified one clone containing cDNA that hybridized to mRNA coding for a M(r) 31,000 IgE-binding protein. The DNA sequence of this cloned cDNA (571 base pairs) was determined and the amino acid sequence corresponding to part of the protein was deduced. In RNA blot analysis, the cDNA hybridized with a mRNA of 1100 nucleotides found in RBL cells but absent in cells not expressing IgE receptors. This cloned cDNA most likely codes for the M(r) 31,000 IgE-binding protein identified in RBL cells, which appears to be related to the IgE-binding phenotype of the cells and which may have a significant role in the IgE-mediated activation of basophils and mast cells.

AB - The synthesis and function of IgE are dependent on IgE-binding proteins, which include cell surface IgE receptors and IgE-binding lymphokines. To further our understanding of the IgE system, we have engaged in the molecular cloning of genes for some of these proteins. In studying the in vitro translation products of mRNA from rat basophilic leukemia (RBL) cells, we have identified a M(r) 31,000 polypeptide that binds IgE and is also reactive with antibodies to proteins affinity-purified from RBL cells with IgE immunoadsorbent. For the molecular cloning, double-stranded cDNA was synthesized from sucrose gradient-fractionated RBL mRNA, inserted into plasmid pBR322, and used to transform Escherichia coli. By screening transformants with a hybridization-selection/in vitro translation procedure, we identified one clone containing cDNA that hybridized to mRNA coding for a M(r) 31,000 IgE-binding protein. The DNA sequence of this cloned cDNA (571 base pairs) was determined and the amino acid sequence corresponding to part of the protein was deduced. In RNA blot analysis, the cDNA hybridized with a mRNA of 1100 nucleotides found in RBL cells but absent in cells not expressing IgE receptors. This cloned cDNA most likely codes for the M(r) 31,000 IgE-binding protein identified in RBL cells, which appears to be related to the IgE-binding phenotype of the cells and which may have a significant role in the IgE-mediated activation of basophils and mast cells.

UR - http://www.scopus.com/inward/record.url?scp=0021821186&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021821186&partnerID=8YFLogxK

U2 - 10.1073/pnas.82.12.4100

DO - 10.1073/pnas.82.12.4100

M3 - Article

VL - 82

SP - 4100

EP - 4104

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 12

ER -