Mechanistic explanations for counter‐intuitive phosphorylation dynamics of the insulin receptor and insulin receptor substrate‐1 in response to insulin in murine adipocytes

• Published in: The FEBS journal 2012-03, Vol.279 (6), p.987-999
• Main Authors: Nyman, Elin, Fagerholm, Siri, Jullesson, David, Strålfors, Peter, Cedersund, Gunnar
• Format: Article
• Language: English
• Subjects: Adipocytes - metabolism; Animals; Body fat; Cell Membrane - metabolism; Cells, Cultured; Cellular biology; conclusive mathematical modeling; core prediction; Insulin; Insulin - metabolism; Insulin Receptor Substrate Proteins - chemistry; Insulin Receptor Substrate Proteins - metabolism; insulin signaling; Kinetics; mechanistic explanation; MEDICIN; MEDICINE; Models, Biological; Phosphorylation; rat adipocytes; Rats; Rats, Sprague-Dawley; Receptor, Insulin - chemistry; Receptor, Insulin - metabolism; Signal Transduction
• ISSN: 1742-464X; 1742-4658; 1742-4658
• DOI: 10.1111/j.1742-4658.2012.08488.x

Insulin signaling through insulin receptor (IR) and insulin receptor substrate‐1 (IRS1) is important for insulin control of target cells. We have previously demonstrated a rapid and simultaneous overshoot behavior in the phosphorylation dynamics of IR and IRS1 in human adipocytes. Herein, we demonstrate that in murine adipocytes a similar overshoot behavior is not simultaneous for IR and IRS1. The peak of IRS1 phosphorylation, which is a direct consequence of the phosphorylation and the activation of IR, occurs earlier than the peak of IR phosphorylation. We used a conclusive modeling framework to unravel the mechanisms behind this counter‐intuitive order of phosphorylation. Through a number of rejections, we demonstrate that two fundamentally different mechanisms may create the reversed order of peaks: (i) two pools of phosphorylated IR, where a large pool of internalized IR peaks late, but phosphorylation of IRS1 is governed by a small plasma membrane‐localized pool of IR with an early peak, or (ii) inhibition of the IR‐catalyzed phosphorylation of IRS1 by negative feedback. Although (i) may explain the reversed order, this two‐pool hypothesis alone requires extensive internalization of IR, which is not supported by experimental data. However, with the additional assumption of limiting concentrations of IRS1, (i) can explain all data. Also, (ii) can explain all available data. Our findings illustrate how modeling can potentiate reasoning, to help draw nontrivial conclusions regarding competing mechanisms in signaling networks. Our work also reveals new differences between human and murine insulin signaling. Database 
The mathematical model described here has been submitted to the Online Cellular Systems Modelling Database and can be accessed at http://jjj.biochem.sun.ac.za/database/nyman/index.html free of charge. We report a counter‐intuitive reversed order of peak values of protein phosphorylation in the early insulin signaling events in murine adipocytes. Using a conclusive modeling framework, we test different hypotheses and are able to draw non‐trivial conclusions regarding possible mechanisms behind the reversed order of peaks. We also identify new differences between human and murine insulin signaling.