Functional reconstitution of insulin receptor binding site from non-binding receptor fragments.

The Journal of Biological Chemistry
Claus KristensenJ Brandt

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
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
Apr 10, 1999·Journal of Structural Biology·P A TullochC W Ward
Aug 14, 1999·Science·R Z LuoF P Ottensmeyer
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

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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
May 22, 2010·Journal of Molecular Biology·Harish Vashisth, Cameron F Abrams
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