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Published on: April 6, 2019
Imidazoline receptor antisera-selected (IRAS) cDNA: cloning and characterization
J E Piletz1, T R Ivanov, J D Sharp
1Department of Psychiatry, University of Mississippi Medical Center, Jackson 39216-4505, USA. jpiletz@psychiatry.umsmed.edu
DNA and Cell Biology
|July 6, 2000
Summary
Researchers identified the first protein with characteristics of an imidazoline-1 receptor (IR1), a novel drug target. This discovery, IRAS-1, advances understanding of IR1 function and potential therapeutic applications.
Area of Science:
- Neuropharmacology
- Molecular Biology
- Drug Discovery
Background:
- The imidazoline-1 receptor (IR1) is a novel target for drug discovery.
- Cloning of IR1 has been challenging due to its unique characteristics.
Purpose of the Study:
- To clone and characterize the imidazoline-1 receptor (IR1).
- To identify the protein responsible for I1 binding sites.
Main Methods:
- Isolation of a truncated cDNA clone from a human hippocampal library.
- 5' and 3' end amplification, end-to-end PCR, and conventional cloning.
- Expression of the IRAS cDNA in Chinese hamster ovary cells and characterization of binding properties.
Main Results:
- A 5131-basepair cDNA, IRAS, encoding a 1504-amino acid protein (IRAS-1) was identified.
- IRAS-1 sequence showed no similarity to known imidazoline-binding proteins but contained cytokine receptor signaling motifs.
- Ruthenium red stained IRAS-1 and inhibited radioligand binding to native I1 sites.
- Transfection of IRAS cDNA into CHO cells created high-affinity I1 binding sites.
Conclusions:
- IRAS-1 is the first identified protein exhibiting characteristics of an imidazoline-1 receptor (IR1).
- This discovery provides a molecular basis for I1 binding sites and opens new avenues for drug development targeting IR1.
