Peptide-based radioimmunoassay for the two isoforms of the human insulin receptor

G. Sesti, R. D'Alfonso, M. D. Vargas Punti, L. Frittitta, V. Trischitta, Y. Y. Liu, P. Borboni, R. Longhi, A. Montemurro, R. Lauro

Research output: Contribution to journalArticlepeer-review

Abstract

The insulin receptor exists in two isoforms differing by the absence (HIR-A) or presence (HIR-B) of 12 amino acids in the COOH-terminus of the α-subunit as a consequence of alternative splicing of exon 11. In this study, we developed a radioimmunoassay for the two isoforms employing antibodies raised against two peptides, one (Pep-12) corresponding to residues encoded by exon 11, and the other (Pep-13) corresponding to a COOH-terminal domain of the α-subunit which is common to both HIR-A and HIR-B isoforms. These peptides were iodinated and used as both ligands and standards. The assay is specific, highly reproducible, and sensitive with a detection limit of 10 fmol of receptor. One mole of purified insulin receptor, measured by Scatchard analysis, is read as one mole of receptor in the radioimmunoassay with either Pep-12 or Pep-13 as standards. The radioimmunoassay is applicable to the measurement of total content and relative abundance of the two isoforms in extracts from various tissues. We applied the radioimmunoassay to measure the relative abundance of the two isoforms in fat and muscle from normal, obese non-diabetic and non-insulin-dependent diabetic (NIDDM) subjects. Results demonstrate that expression of the low-affinity HIR-B form is significantly increased in obese and NIDDM subjects compared with control subjects. In addition, the increased expression of the HIR-B isoform was significantly correlated with both body mass index (r=0.52; p=0.006) and fasting glucose levels (r=0.59; p=0.001).

Original languageEnglish
Pages (from-to)445-453
Number of pages9
JournalDiabetologia
Volume38
Issue number4
DOIs
Publication statusPublished - Apr 1995

Keywords

  • Insulin receptor isoforms
  • insulin receptor radioimmunoassay

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Fingerprint Dive into the research topics of 'Peptide-based radioimmunoassay for the two isoforms of the human insulin receptor'. Together they form a unique fingerprint.

Cite this