Nucleotide and amino acid sequence of lymphocyte-derived corticotropin

Endotoxin induction of a truncated peptide

Eric M. Smith, F. Shawn Galin, Robert D. LeBoeuf, Dorian H. Coppenhaver, Deborah V. Harbour, J. Edwin Blalock

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

To determine the degree of similarity between pituitary and lymphocyte proopiomelanocortin, the lymphocyte mRNA was reverse transcribed, cloned, and sequenced. Murine lymphocyte mRNA was first purified by oligo(dT)-cellulose affinity chromatography and was reverse transcribed by using a selective 3′ antisense oligonucleotide primer directed at the boundary between the translated/nontranslated region on the 3′ end of exon 3. This cDNA was then amplified in a polymerase chain reaction with selective primers containing Sal I and Kpn I restriction endonuclease sites. Amplified cDNA was then directionally ligated into M13mp18 and M13mp19 bacteriophage and was sequenced. The nucleotide sequence encoding this peptide was identical to that of mouse pituitary corticotropin (ACTH). Elevated levels of lymphocyte immunoreactive ACTH were then induced with bacterial lipopolysaccharide and the peptide(s) was purified by antibody affinity chromatography and reverse-phase high-performance liquid chromatography. The predominant immunoreactive ACTH species was ≈3 kDa and its sequence was identical to pituitary ACTH(1-25). These results conclusively demonstrate that lymphocytes produce authentic ACTH and harbor its mRNA.

Original languageEnglish (US)
Pages (from-to)1057-1060
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number3
StatePublished - 1990

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Endotoxins
Adrenocorticotropic Hormone
Amino Acid Sequence
Nucleotides
Lymphocytes
Peptides
Affinity Chromatography
Messenger RNA
Complementary DNA
Pro-Opiomelanocortin
Antibody Affinity
DNA Primers
Antisense Oligonucleotides
DNA Restriction Enzymes
Reverse-Phase Chromatography
Bacteriophages
Lipopolysaccharides
Exons
High Pressure Liquid Chromatography
Polymerase Chain Reaction

Keywords

  • Immune neuroendocrine interactions
  • Protein microsequencing

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Nucleotide and amino acid sequence of lymphocyte-derived corticotropin : Endotoxin induction of a truncated peptide. / Smith, Eric M.; Galin, F. Shawn; LeBoeuf, Robert D.; Coppenhaver, Dorian H.; Harbour, Deborah V.; Blalock, J. Edwin.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 3, 1990, p. 1057-1060.

Research output: Contribution to journalArticle

Smith, Eric M. ; Galin, F. Shawn ; LeBoeuf, Robert D. ; Coppenhaver, Dorian H. ; Harbour, Deborah V. ; Blalock, J. Edwin. / Nucleotide and amino acid sequence of lymphocyte-derived corticotropin : Endotoxin induction of a truncated peptide. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 3. pp. 1057-1060.
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