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Involvement of non‐conserved residues important for PGE 2 binding to the constrained EP3 eLP 2 using NMR and site‐directed mutagenesis
Author(s) -
Chillar Annirudha,
Wu Jiaxin,
So Shui-Ping,
Ruan Ke-He
Publication year - 2008
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2008.07.018
Subject(s) - mutagenesis , site directed mutagenesis , chemistry , binding site , stereochemistry , computational biology , biochemistry , mutation , biology , mutant , gene
A peptide constrained to a conformation of second extracellular loop of human prostaglandin‐E 2 (PGE 2 ) receptor subtype3 (hEP3) was synthesized. The contacts between the peptide residues at S211 and R214, and PGE 2 were first identified by NMR spectroscopy. The results were used as a guide for site‐directed mutagenesis of the hEP3 protein. The S211L and R214L mutants expressed in HEK293 cells lost binding to [ 3 H]PGE 2 . This study found that the non‐conserved S211 and R214 of the hEP3 are involved in PGE 2 recognition, and implied that the corresponding residues in other subtype receptors could be important to distinguish the different configurations of PGE 2 ligand recognition sites.