PMID: 2500341Jun 15, 1989Paper

Antibodies directed against phosphothreonine residues as potent tools for studying protein phosphorylation

European Journal of Biochemistry
D HeffetzY Zick

Abstract

Here we report the development of novel antibodies which specifically react with phosphothreonine residues [anti-(P-Thr)antibodies]. The specificity of the antibodies was assessed in radioimmunoassays where we could demonstrate that half-maximal and maximal binding of the antibodies to plates coated with BSA - P-Thr occurred at serum dilutions of 1:4000 and 1:1000, respectively. P-Thr inhibited antibody binding with a half-maximal effect at 40 microM. P-Ser was 200-fold less potent while P-Tyr was essentially ineffective. Anti-(P-Thr) antibodies could specifically bind to phosphothreonine-containing proteins on Western blots. Using such a procedure we could demonstrate enhanced threonine phosphorylation of the EGF receptor upon treatment of intact unlabeled A431 cells with EGF. We could further demonstrate antibodies binding to proteins present in extracts of rat hepatoma cells (Fao). P-Thr at 10 microM completely inhibited antibody binding while P-Ser, P-Tyr, Thr or Ser, each present at tenfold higher concentrations, had no such inhibitory effect. Anti-(P-Thr) antibodies were also capable of specifically immunoprecipitating 32P-labeled phosphoproteins present in Triton extracts of Fao cells. Immunoprecipitation of proteins of ...Continue Reading

References

Sep 11, 1987·Cell·T Hunter
Nov 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·A O Morla, J Y Wang
Jan 1, 1983·Methods in Enzymology·J A CooperT Hunter

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Citations

Sep 25, 2008·Proteomics·Poh-Kuan ChongYoon-Pin Lim
Oct 13, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Stefan R SchmidtMartin E Andersson
Jan 30, 2008·Current Protocols in Cell Biology·Z Yao, R Seger
Jul 6, 2011·Journal of Separation Science·Zdenka KučerováMarie Tichá
May 25, 2018·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·C SidhanthT S Ganesan
May 1, 1992·Journal of Pharmaceutical and Biomedical Analysis·H KataokaM Makita
Nov 20, 2001·Chemical Reviews·R Aebersold, D R Goodlett

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