Purification of the human blood platelet thromboxane A2/prostaglandin H2 receptor protein

Biochemical Pharmacology
S O KimG C Le Breton

Abstract

The human platelet thromboxane A2/prostaglandin H2 receptor has been purified 6100-fold to apparent homogeneity by a three-step chromatographic procedure with an overall yield of 6%. A 6-fold purification of the receptor was first achieved by chromatography of 3-[(3-cholamidopropyl)dimethyl-ammonio]-1-propanesulfonate (CHAPS)-solubilized membrane proteins from human platelets on a diethylaminoethyl (DEAE)-Sepharose column. The DEAE eluate fractions containing receptor activity were then applied to a newly developed affinity column using the cyclohexyl derivative of SQ30,741 (SQ31,491) as the immobilized ligand. Elution of the receptor from the affinity column with BM13.177 yielded a further purification of 1700-fold. An additional 4-fold receptor purification from the affinity column eluate was achieved by HPLC using GPC 500 and GPC 100 columns connected in tandem. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining of the HPLC eluate containing purified receptor revealed a single, distinct band with a molecular weight of 55,000. The receptor binding activity was detected with [3H]SQ29,548 using a newly developed binding assay which involved immobilization of the receptor on polyethyleneimine-treated g...Continue Reading

References

Jul 1, 1979·Journal of Medicinal Chemistry·D L VentonG C Le Breton
Aug 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·G C Le BretonP V Halushka
Sep 1, 1977·Prostaglandins·J SvenssenM Hamberg
Aug 30, 1989·Biochemical and Biophysical Research Communications·G W Dorn
Nov 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·R M BurchP V Halushka
Aug 1, 1983·British Journal of Pharmacology·R A ArmstrongN H Wilson
May 15, 1983·Thrombosis Research·M Radomski, S Moncada

Citations

Sep 2, 1998·Proceedings of the National Academy of Sciences of the United States of America·Y DjellasG C Le Breton
May 1, 1996·Kidney International·B A BresnahanE A Lianos
Sep 23, 2010·European Journal of Pharmacology·Harold J Ting, Fadi T Khasawneh
Jan 4, 2012·Journal of Thrombosis and Haemostasis : JTH·Y ZhangY Zhang
Apr 29, 2015·PloS One·Matthew F KeeYongzhi Qiu
Feb 5, 2014·British Journal of Pharmacology·K T SanthoshS Dakshinamurti
May 13, 1999·The Journal of Biological Chemistry·Y DjellasG C Le Breton
Sep 17, 1999·The Journal of Biological Chemistry·J M ManganelloG C Le Breton
Feb 7, 1998·The Journal of Biological Chemistry·S C BlackmanG C Le Breton
Aug 21, 2015·The Journal of Biological Chemistry·Subhashini SrinivasanG C Le Breton
Nov 9, 2011·Journal of Cardiovascular Pharmacology and Therapeutics·Harold J TingFadi T Khasawneh
Jan 1, 1994·Journal of Ocular Pharmacology·N Sano, H Shichi

Related Concepts

7-(3-((2-((phenylamino)carbonyl)hydrazino)methyl)-7-oxabicyclo(2.2.1)hept-2-yl)-5-heptenoic acid
3-((((-oxoheptyl)amino)acetyl)amino)methyl-7-oxobicyclo(2.2.1)hept-2-yl-5-heptenoic acid
Blood Platelets
Chromatography, Affinity
High Pressure Liquid Chromatography Procedure
Ion-Exchange Chromatography Procedure
SDS-PAGE
Fatty Acids, Unsaturated
Hydrazines
Cell Surface Proteins

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