Reactivity of thyrotropin receptor autoantibodies with the thyrotropin receptor on western blots

Thyroid : Official Journal of the American Thyroid Association
M MaruyamaB Rees Smith

Abstract

Affinity purified recombinant human thyrotropin receptor (TSHR) was run on sodium dodecyl sulfate (SDS) gels and subjected to a renaturing and blotting procedure. Twenty sera from thyrotropin receptor autoantibodies (TRAb)-positive patients with a history of hyperthyroidism and 20 sera with high levels of TSH blocking activity were analyzed. Four of 20 sera with blocking-type of TRAb (i.e., TSH antagonist activity) were able to recognize the mature, fully glycosylated 120-kd form of the receptor on blots of gels run under reducing conditions. No sera recognized the 100-kd high mannose precursor form of the TSHR. Three of the four recognized a 74-kd band and 2 of the 4 recognized a 50-kd band. These bands are probably proteolytic cleavage fragments of the mature 120-kd TSHR. In the absence of reducing agent the same 4 of 20 sera described above together with a further serum sample (i.e., 5/20 in total) reacted with the 120-kd form of the receptor. No specific reaction with the TSHR was observed on Western blots with the remaining 15 sera with TSH blocking activity, nor with 20 sera from patients with a history of hyperthyroidism, nor with sera from 10 healthy blood donors, 10 Hashimoto sera (negative for TRAb) and 10 systemic lu...Continue Reading

References

Feb 1, 1988·Endocrine Reviews·B Rees SmithJ Furmaniak
May 1, 1984·Clinical Endocrinology·K SouthgateB Rees Smith
Aug 14, 1996·Biochemical and Biophysical Research Communications·G D ChazenbalkB Rapoport
May 19, 1998·Journal of Molecular Endocrinology·Y OdaB Rees Smith
Oct 16, 1999·European Journal of Biochemistry·M AtgerE Milgrom
Feb 24, 2001·Thyroid : Official Journal of the American Thyroid Association·Y OdaB R Smith

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Citations

Apr 14, 2005·Best Practice & Research. Clinical Endocrinology & Metabolism·Takao AndoTerry F Davies
Aug 21, 2002·Thyroid : Official Journal of the American Thyroid Association·T Akamizu
Feb 21, 2003·Thyroid : Official Journal of the American Thyroid Association·Jane SandersBernard Rees Smith
Feb 21, 2003·Thyroid : Official Journal of the American Thyroid Association·Jennifer JeffreysBernard Rees Smith
Aug 2, 2005·Thyroid : Official Journal of the American Thyroid Association·J SandersB Rees Smith
Jul 18, 2008·Thyroid : Official Journal of the American Thyroid Association·Jane SandersBernard Rees Smith
Jul 18, 2002·The Journal of Clinical Investigation·Gregorio D ChazenbalkBasil Rapoport
Jul 30, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Rachel HewerBernard Rees Smith
Jul 3, 2010·Autoimmunity Reviews·Klaus ZöphelMatthias Schott

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