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Synthesis and processing of D2 dopamine receptors
C David1, C S Fishburn, F J Monsma
1Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Biochemistry
|August 17, 1993
Summary
Researchers visualized dopamine receptor synthesis using Chinese hamster ovary cells. A 35-kDa precursor protein is processed into a 75-kDa glycosylated dopamine receptor.
Area of Science:
- Neuroscience
- Molecular Biology
Background:
- Dopamine receptors are neurotransmitter receptors coupled to guanine nucleotide binding proteins.
- Understanding dopamine receptor synthesis is crucial for neuroscience research.
Purpose of the Study:
- To visualize the synthesis and processing of the rat D2L dopamine receptor.
- To identify precursor forms of dopamine receptors.
Main Methods:
- Utilized Chinese hamster ovary (CHO) cells stably transfected with the rat D2L receptor.
- Employed specific anti-peptide antibodies for protein visualization.
- Conducted pulse-chase experiments and in vitro translation systems.
Main Results:
- Newly synthesized receptors appear as a 45-kDa protein, processed to a 75-kDa glycosylated form in CHO cells.
- A 35-kDa precursor protein was observed, disappearing within 30 minutes.
- In vitro translation of murine D2 and D3 receptor cDNAs yielded 35-kDa proteins, which could be glycosylated to ~45 kDa.
Conclusions:
- The 35-kDa protein represents an unprocessed form of the dopamine receptor.
- Dopamine receptors undergo significant post-translational modification, including glycosylation.