Abstract
We have previously shown that a minimized insulin receptor (IR) consisting of the first 468 amino acids of the insulin receptor fused to 16 amino acids from the C terminus of the alpha-subunit (CT domain) bound insulin with nanomolar affinity (Kristensen, C., Wiberg, F. C., Schäffer, L., and Andersen, A. S. (1998) J. Biol. Chem. 273, 17780-17786). In the present study, we show that a smaller construct that has the first 308 residues fused to the CT domain also binds insulin. Insulin receptor fragments consisting of the first 468 or 308 residues did not bind insulin. However, when these fragments were mixed with a synthetic peptide corresponding to the CT domain, insulin binding was detectable. At concentrations of 10 microm CT peptide, insulin binding was fully reconstituted yielding apparent affinities of 9-11 nm. To further investigate the minimum requirement for the length of the N terminus of IR, we tested smaller receptor fragments for insulin binding in the presence of the CT peptide and found that a fragment consisting of the first 255 amino acids of IR was able to fully reconstitute the insulin binding site, yielding an apparent affinity of 11 +/- 4 nm for insulin.
References
Sep 1, 1992·Biological Chemistry Hoppe-Seyler·M Fabry, D Brandenburg
Aug 14, 1990·Biochemistry·A S AndersenN P Møller
Mar 21, 1989·Biochemistry·S M Waugh, P F Pilch
Feb 6, 1985·Nature·A UllrichM Tsubokawa
Nov 26, 1987·Biochimica Et Biophysica Acta·M BajajT Blundell
Jun 1, 1995·Proteins·C W WardR H Flegg
Sep 1, 1994·Diabetologia·P De Meyts
Sep 1, 1994·Diabetes·P RoachP Gorden
May 1, 1994·European Journal of Biochemistry·L Schäffer
Feb 2, 1996·The Journal of Biological Chemistry·D C MynarcikJ Whittaker
Dec 31, 1997·The Journal of Biological Chemistry·L G SparrowC W Ward
Jul 4, 1998·The Journal of Biological Chemistry·C KristensenA S Andersen
Aug 5, 1998·Nature·T P GarrettC W Ward
Apr 10, 1999·Journal of Structural Biology·P A TullochC W Ward
Aug 14, 1999·Science·R Z LuoF P Ottensmeyer
Dec 22, 1999·The Journal of Biological Chemistry·C KristensenA S Andersen
Mar 10, 2000·FEBS Letters·P A HoyneC W Ward
Aug 5, 2000·Biochemical Society Transactions·C Marino-BusljeT L Blundell
Aug 29, 2000·Cellular and Molecular Life Sciences : CMLS·T E AdamsC W Ward
Mar 30, 2001·The Journal of Biological Chemistry·J BrandtC Kristensen
Aug 14, 2001·The Journal of Biological Chemistry·J WhittakerL J Whittaker
Mar 5, 2002·The Journal of Biological Chemistry·Katharina Helen SurinyaKenneth Siddle
Citations
Oct 3, 2002·Nature Reviews. Drug Discovery·Pierre De Meyts, Jonathan Whittaker
May 27, 2008·The Journal of Biological Chemistry·Lisbeth GauguinPierre De Meyts
Jun 1, 2011·PloS One·Tine GlendorfThomas Kjeldsen
Aug 9, 2006·Proceedings of the National Academy of Sciences of the United States of America·Meizhen LouColin W Ward
Aug 6, 2014·Proceedings of the National Academy of Sciences of the United States of America·John G MentingMichael C Lawrence
Oct 14, 2014·Membranes·Harish Vashisth
Feb 14, 2015·Membranes·Harish Vashisth
May 22, 2010·Journal of Molecular Biology·Harish Vashisth, Cameron F Abrams
Jul 22, 2008·Trends in Biochemical Sciences·Pierre De Meyts
Feb 20, 2009·Molecular Systems Biology·Vladislav V KiselyovPierre De Meyts
Jan 26, 2013·Proteins·Harish Vashisth, Cameron F Abrams
Nov 28, 2013·Journal of Molecular Recognition : JMR·Kannan SubramanianMark T Hayes
Jan 30, 2010·Proteins·Harish Vashisth, Cameron F Abrams
Jul 4, 2006·Biochemical and Biophysical Research Communications·Caili HaoJonathan Whittaker
May 4, 2016·Journal of Biomolecular Structure & Dynamics·Kei C SitTristan Croll
Nov 23, 2016·The Journal of Biological Chemistry·Krystel El HageMichael A Weiss
Sep 20, 2018·Diabetes, Obesity & Metabolism·Michael A Weiss, Michael C Lawrence
Feb 28, 2018·Nature Communications·Yibin XuMichael C Lawrence
Jan 23, 2003·Endocrinology·S GeorgeS O'Rahilly
Jun 19, 2004·Molecular Endocrinology·Adam DenleyJohn C Wallace
Jun 3, 2020·Nature Structural & Molecular Biology·Xiaochun XiongDanny Hung-Chieh Chou
Feb 13, 2007·The Biochemical Journal·Samira BenyoucefKenneth Siddle
Sep 25, 2002·The Journal of Biological Chemistry·Jonathan WhittakerJane Hinrichsen
Jun 17, 2020·IScience·Jia XieRichard A Lerner
May 23, 2009·Biochemistry·John G MentingMichael C Lawrence
Apr 28, 2006·Journal of Medicinal Chemistry·Yulin RenXiaozhuo Chen
May 17, 2021·Molecular Metabolism·Michael C Lawrence
Dec 22, 2004·Biochemistry·Zhuli WanMichael A Weiss