Purification, cloning, and tissue distribution of a 23-kDa rat protein isolated by morphine affinity chromatography

David Grandy, Eric Hanneman, James Bunzow, Marjorie Shih, Curtis Machida, Jean M. Bidlack, Olivier Civelli

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

A 23-kDa (p23k) rat brain protein was stereospecifically eluted from a 14β-bromoacetamidomorphine affinity column, purified to apparent homogeneity by reverse phase HPLC, and partially sequenced. Three degenerate oligodeoxynucleotide probes were synthesized based on this partial amino acid sequence. A rat brain cDNA library was screened using these probes, and a full-length cDNA was isolated. The deduced protein, 187 amino acids long, is rich in glutamic and aspartic acid residues, endowing p23k with a net negative charge at neutral pH. The protein lacks a signal sequence as well as any transmembrane domains. Based on predictions of secondary structure, p23k is a globular protein composed of 30% α-helices and 18% β-pleated sheets. Northern blot analysis revealed p23k transcripts in rat brain, liver, and the mouse x rat neuroblastoma-glioma NG108-14 cell line. Although not an opioid receptor itself, this protein may be associated with such a receptor or be related to a protein that has been shown to be cross-linked to the opioid peptide β-endorphin.

Original languageEnglish (US)
Pages (from-to)1370-1376
Number of pages7
JournalMolecular Endocrinology
Volume4
Issue number9
StatePublished - 1990

Fingerprint

Tissue Distribution
Affinity Chromatography
Morphine
Organism Cloning
Proteins
Brain
Endorphins
Opioid Peptides
Oligodeoxyribonucleotides
Opioid Receptors
Protein Sorting Signals
Neuroblastoma
Gene Library
Glioma
Northern Blotting
Glutamic Acid
Amino Acid Sequence
Complementary DNA
High Pressure Liquid Chromatography
Amino Acids

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Grandy, D., Hanneman, E., Bunzow, J., Shih, M., Machida, C., Bidlack, J. M., & Civelli, O. (1990). Purification, cloning, and tissue distribution of a 23-kDa rat protein isolated by morphine affinity chromatography. Molecular Endocrinology, 4(9), 1370-1376.

Purification, cloning, and tissue distribution of a 23-kDa rat protein isolated by morphine affinity chromatography. / Grandy, David; Hanneman, Eric; Bunzow, James; Shih, Marjorie; Machida, Curtis; Bidlack, Jean M.; Civelli, Olivier.

In: Molecular Endocrinology, Vol. 4, No. 9, 1990, p. 1370-1376.

Research output: Contribution to journalArticle

Grandy, D, Hanneman, E, Bunzow, J, Shih, M, Machida, C, Bidlack, JM & Civelli, O 1990, 'Purification, cloning, and tissue distribution of a 23-kDa rat protein isolated by morphine affinity chromatography', Molecular Endocrinology, vol. 4, no. 9, pp. 1370-1376.
Grandy, David ; Hanneman, Eric ; Bunzow, James ; Shih, Marjorie ; Machida, Curtis ; Bidlack, Jean M. ; Civelli, Olivier. / Purification, cloning, and tissue distribution of a 23-kDa rat protein isolated by morphine affinity chromatography. In: Molecular Endocrinology. 1990 ; Vol. 4, No. 9. pp. 1370-1376.
@article{a269b4df6c6b4590a0dad00d03cfcc26,
title = "Purification, cloning, and tissue distribution of a 23-kDa rat protein isolated by morphine affinity chromatography",
abstract = "A 23-kDa (p23k) rat brain protein was stereospecifically eluted from a 14β-bromoacetamidomorphine affinity column, purified to apparent homogeneity by reverse phase HPLC, and partially sequenced. Three degenerate oligodeoxynucleotide probes were synthesized based on this partial amino acid sequence. A rat brain cDNA library was screened using these probes, and a full-length cDNA was isolated. The deduced protein, 187 amino acids long, is rich in glutamic and aspartic acid residues, endowing p23k with a net negative charge at neutral pH. The protein lacks a signal sequence as well as any transmembrane domains. Based on predictions of secondary structure, p23k is a globular protein composed of 30{\%} α-helices and 18{\%} β-pleated sheets. Northern blot analysis revealed p23k transcripts in rat brain, liver, and the mouse x rat neuroblastoma-glioma NG108-14 cell line. Although not an opioid receptor itself, this protein may be associated with such a receptor or be related to a protein that has been shown to be cross-linked to the opioid peptide β-endorphin.",
author = "David Grandy and Eric Hanneman and James Bunzow and Marjorie Shih and Curtis Machida and Bidlack, {Jean M.} and Olivier Civelli",
year = "1990",
language = "English (US)",
volume = "4",
pages = "1370--1376",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "9",

}

TY - JOUR

T1 - Purification, cloning, and tissue distribution of a 23-kDa rat protein isolated by morphine affinity chromatography

AU - Grandy, David

AU - Hanneman, Eric

AU - Bunzow, James

AU - Shih, Marjorie

AU - Machida, Curtis

AU - Bidlack, Jean M.

AU - Civelli, Olivier

PY - 1990

Y1 - 1990

N2 - A 23-kDa (p23k) rat brain protein was stereospecifically eluted from a 14β-bromoacetamidomorphine affinity column, purified to apparent homogeneity by reverse phase HPLC, and partially sequenced. Three degenerate oligodeoxynucleotide probes were synthesized based on this partial amino acid sequence. A rat brain cDNA library was screened using these probes, and a full-length cDNA was isolated. The deduced protein, 187 amino acids long, is rich in glutamic and aspartic acid residues, endowing p23k with a net negative charge at neutral pH. The protein lacks a signal sequence as well as any transmembrane domains. Based on predictions of secondary structure, p23k is a globular protein composed of 30% α-helices and 18% β-pleated sheets. Northern blot analysis revealed p23k transcripts in rat brain, liver, and the mouse x rat neuroblastoma-glioma NG108-14 cell line. Although not an opioid receptor itself, this protein may be associated with such a receptor or be related to a protein that has been shown to be cross-linked to the opioid peptide β-endorphin.

AB - A 23-kDa (p23k) rat brain protein was stereospecifically eluted from a 14β-bromoacetamidomorphine affinity column, purified to apparent homogeneity by reverse phase HPLC, and partially sequenced. Three degenerate oligodeoxynucleotide probes were synthesized based on this partial amino acid sequence. A rat brain cDNA library was screened using these probes, and a full-length cDNA was isolated. The deduced protein, 187 amino acids long, is rich in glutamic and aspartic acid residues, endowing p23k with a net negative charge at neutral pH. The protein lacks a signal sequence as well as any transmembrane domains. Based on predictions of secondary structure, p23k is a globular protein composed of 30% α-helices and 18% β-pleated sheets. Northern blot analysis revealed p23k transcripts in rat brain, liver, and the mouse x rat neuroblastoma-glioma NG108-14 cell line. Although not an opioid receptor itself, this protein may be associated with such a receptor or be related to a protein that has been shown to be cross-linked to the opioid peptide β-endorphin.

UR - http://www.scopus.com/inward/record.url?scp=0025084434&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025084434&partnerID=8YFLogxK

M3 - Article

VL - 4

SP - 1370

EP - 1376

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 9

ER -