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 alpha-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 alpha-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).

Sesti, G., D'Alfonso, R., Vargas Punti, M., Frittitta, L., Trischitta, V., Liu, Y., et al. (1995). Peptide-based radioimmunoassay for the two isoforms of the human insulin receptor. DIABETOLOGIA, 38(4), 445-453.

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

SESTI, GIORGIO;D'ALFONSO, ROSSELLA;BORBONI, PATRIZIA;MONTEMURRO, ANTONIO;LAURO, RENATO
1995-04-01

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 alpha-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 alpha-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).
apr-1995
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore MED/09 - MEDICINA INTERNA
Settore MED/13 - ENDOCRINOLOGIA
Settore BIO/12 - BIOCHIMICA CLINICA E BIOLOGIA MOLECOLARE CLINICA
English
Con Impact Factor ISI
3T3 Cells; Placenta; Middle Aged; Radioimmunoassay; Transfection; Binding, Competitive; Diabetes Mellitus, Type 2; Alternative Splicing; Obesity; Receptor, Insulin; Mice; Pregnancy; Recombinant Proteins; Adipose Tissue; Reference Values; Antibody Specificity; Muscle, Skeletal; Male; Organ Specificity; Female; Antibodies; Animals; Humans; Kinetics; Regression Analysis; Peptide Fragments; Liver; Exons
Sesti, G., D'Alfonso, R., Vargas Punti, M., Frittitta, L., Trischitta, V., Liu, Y., et al. (1995). Peptide-based radioimmunoassay for the two isoforms of the human insulin receptor. DIABETOLOGIA, 38(4), 445-453.
Sesti, G; D'Alfonso, R; Vargas Punti, M; Frittitta, L; Trischitta, V; Liu, Y; Borboni, P; Longhi, R; Montemurro, A; Lauro, R
Articolo su rivista
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/54615
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 26
  • ???jsp.display-item.citation.isi??? ND
social impact