Generation of met-enkephalin arg6phe7 immunoreactivity by proteolytic cleavage of mammalian plasma precursors by pepsin

Andrew S. Giraud, Lorraine Parker, Coral Reichman, Mary Familari, A. Ian Smith, John Funder

Research output: Contribution to journalArticleResearchpeer-review

Abstract

A region-specific antiserum raised against the C-terminal heptapeptide of proenkephalin A (Met-enk Arg6Phe7) was used in RIA studies to show that rat, human, and ovine plasma contain substrates (mol wt, 68K) that yield nanomolar amounts of Met-enk Arg6Phe7 (ME-RF) after treatment with pepsin under acid conditions. This ovine plasma-derived immunoreactivity diluted in parallel to the ME-RF standard in RIA and chromatographed as two low mol wt species (∼ IK) which were less hydrophobic than the standard on size exclusion and reverse phase chromatography. The pepsin-generated material displaced [3H]naloxone from rat brain binding sites; its potency was about 1000-fold that of ME-RF, assuming near 100% cross-reactivity with the antiserum. Taken together these observations suggest that the pepsin-generated material is of similar mol wt and amino acid sequence to ME-RF, but differs with respect to opiate-binding efficacy, and that the plasma precursor is distinct from proenkephalin in both size and processing sites.

Original languageEnglish
Pages (from-to)1711-1716
Number of pages6
JournalEndocrinology
Volume124
Issue number4
DOIs
Publication statusPublished - 1 Jan 1989
Externally publishedYes

Cite this

Giraud, Andrew S. ; Parker, Lorraine ; Reichman, Coral ; Familari, Mary ; Smith, A. Ian ; Funder, John. / Generation of met-enkephalin arg6phe7 immunoreactivity by proteolytic cleavage of mammalian plasma precursors by pepsin. In: Endocrinology. 1989 ; Vol. 124, No. 4. pp. 1711-1716.
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abstract = "A region-specific antiserum raised against the C-terminal heptapeptide of proenkephalin A (Met-enk Arg6Phe7) was used in RIA studies to show that rat, human, and ovine plasma contain substrates (mol wt, 68K) that yield nanomolar amounts of Met-enk Arg6Phe7 (ME-RF) after treatment with pepsin under acid conditions. This ovine plasma-derived immunoreactivity diluted in parallel to the ME-RF standard in RIA and chromatographed as two low mol wt species (∼ IK) which were less hydrophobic than the standard on size exclusion and reverse phase chromatography. The pepsin-generated material displaced [3H]naloxone from rat brain binding sites; its potency was about 1000-fold that of ME-RF, assuming near 100{\%} cross-reactivity with the antiserum. Taken together these observations suggest that the pepsin-generated material is of similar mol wt and amino acid sequence to ME-RF, but differs with respect to opiate-binding efficacy, and that the plasma precursor is distinct from proenkephalin in both size and processing sites.",
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Generation of met-enkephalin arg6phe7 immunoreactivity by proteolytic cleavage of mammalian plasma precursors by pepsin. / Giraud, Andrew S.; Parker, Lorraine; Reichman, Coral; Familari, Mary; Smith, A. Ian; Funder, John.

In: Endocrinology, Vol. 124, No. 4, 01.01.1989, p. 1711-1716.

Research output: Contribution to journalArticleResearchpeer-review

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