High affinity insulin binding in the human placenta insulin receptor requires αβ heterodimeric subunit interactions

Michael L. Swanson, Jeffrey E. Pessin

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Insulin binding to human placenta membranes treated at pH 7.6 or 8.5 in the presence or absence of 2.0 mm DTT for 5 min, followed by the simultaneous removal of the DTT and pH adjustment to pH 7.6, displayed curvilinear (heterogeneous) insulin binding plots when analyzed by the method of Scatchard. However, Triton X-100 solubilization followed by Bio-Gel A-1.5m gel filtration chromatography of the placenta membranes previously treated with DTT at pH 8.5 generated a nearly straight line (homogeneous) Scatchard plot.125I-insulin affinity crosslinking studies coupled with Bio-Gel A-1.5m gel filtration chromatography demonstrated that the alkaline pH and DTT treatment of placenta membranes followed by detergent solubilization generated an αβ heterodimeric insulin receptor complex from the α2β2 heterotetrameric disulfide-linked state. The ability of alkaline pH and DTT to produce a functional αβ heterodimeric insulin receptor complex was found to be time dependent with maximal formation and preservation of tracer insulin binding occurring at 5 min. These data demonstrate that (i) a combination of alkaline pH and DTT treatment of placenta membranes can result in the formation of a functional αβ heterodimeric insulin receptor complex. (ii) the αβ heterodimeric complex displays homogeneous insulin binding. (iii) the insulin receptor membrane environment maintains the α2β2 association state, which displays heterogeneous insulin binding, despite reduction of the critical domains that are responsible for the covalent interaction between the αβ heterodimers.

Original languageEnglish (US)
Pages (from-to)217-225
Number of pages9
JournalThe Journal of Membrane Biology
Volume108
Issue number3
DOIs
StatePublished - Jun 1989
Externally publishedYes

Fingerprint

Placenta
Insulin
Insulin Receptor
Membranes
Gel Chromatography
Gels
Octoxynol
human INSR protein
Disulfides
Detergents

Keywords

  • insulin binding
  • insulin receptor
  • subunit interaction

ASJC Scopus subject areas

  • Physiology
  • Cell Biology
  • Biophysics

Cite this

High affinity insulin binding in the human placenta insulin receptor requires αβ heterodimeric subunit interactions. / Swanson, Michael L.; Pessin, Jeffrey E.

In: The Journal of Membrane Biology, Vol. 108, No. 3, 06.1989, p. 217-225.

Research output: Contribution to journalArticle

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