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Purification of brain D2 dopamine receptor
R A Williamson1, S Worrall, P L Chazot
1Biological Laboratory, The University, Canterbury, Kent, UK.
The EMBO Journal
|December 20, 1988
Summary
Researchers purified D2 dopamine receptors from bovine brain using affinity chromatography. The study identified the D2 dopamine receptor as a glycoprotein with a molecular weight of 95,000.
Area of Science:
- Neuroscience
- Biochemistry
- Molecular Biology
Background:
- D2 dopamine receptors are crucial for neurotransmission.
- Understanding their structure is key to developing targeted therapeutics.
Purpose of the Study:
- To purify and characterize the D2 dopamine receptor.
- To determine the molecular weight and nature of the purified receptor.
Main Methods:
- Bovine brain tissue was used for receptor extraction.
- Detergent cholate and affinity chromatography (haloperidol-sepharose, wheat germ agglutinin-agarose) were employed for purification.
- Pharmacological specificity of [3H]spiperone binding and SDS-polyacrylamide gel electrophoresis were used for characterization.
Main Results:
- D2 dopamine receptors were purified approximately 20,000-fold.
- The purified preparation exhibited specific [3H]spiperone binding, confirming D2 receptor presence.
- SDS-PAGE revealed a major band at Mr 95,000, with evidence of microheterogeneity.
- Photoaffinity labeling also indicated a Mr 95,000 species.
Conclusions:
- The D2 dopamine receptor is a glycoprotein.
- The molecular weight of the D2 dopamine receptor was determined to be 95,000.