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Measurement of T-kinin in rat plasma using a specific radioimmunoassay.

Abstract
T-kinin (Ile-Ser-Bradykinin) has been isolated only from the plasma of the rat and it is unclear whether the peptide, or its biosynthetic precursor, T-kininogen, circulates in the human. An NH2-terminally directed antiserum to T-kinin was raised in rabbits using an immunogen prepared by coupling the free -SH group of T-kinin extended from its COOH-terminus by a cysteinyl residue to an -NH2 group on human serum albumin. A radioimmunoassay was developed using this antiserum and 125I-labelled [Tyr10]T-kinin as tracer that was sensitive (least-detectable concentration 3 fmol/tube) and relatively specific for T-kinin (cross-reactivity with bradykinin and kallidin less than 1%). Treatment of rat plasma with an excess of trypsin in the presence of a kininase inhibitor generated T-kinin immunoreactivity equivalent to 455 +/- 71 pmol/ml (mean +/- S.E.M.; n = 9) and this immunoreactivity was eluted from a reversed-phase HPLC column as a single peak with the same retention time as synthetic T-kinin. In contrast, treatment of plasma from healthy human subjects (n = 8) and from patients (n = 8) with inflammation due to acute or chronic gastrointestinal disease under the same conditions did not generate any detectable T-kinin immunoreactivity. It is concluded, therefore, that T-kininogen, the biosynthetic precursor of T-kinin in the rat, is either absent from the plasma of human subjects or is present in a concentration less than 30 fmol/ml. Similarly, T-kininogen is probably not an acute phase reactant in humans.
AuthorsF O'Harte, D D Smith, S J Lanspa, J M Conlon
JournalRegulatory peptides (Regul Pept) Vol. 41 Issue 2 Pg. 139-48 (Sep 22 1992) ISSN: 0167-0115 [Print] Netherlands
PMID1438985 (Publication Type: Journal Article)
Chemical References
  • Kininogens
  • T-kinin
  • Bradykinin
Topics
  • Adult
  • Amino Acid Sequence
  • Animals
  • Bradykinin (analogs & derivatives, blood, chemistry)
  • Female
  • Gastroenteritis (blood)
  • Humans
  • Kininogens (blood)
  • Male
  • Molecular Sequence Data
  • Radioimmunoassay (methods)
  • Rats
  • Rats, Sprague-Dawley
  • Species Specificity

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