Alanine exchanges of polar amino acids in the transmembrane domains of a platelet-activating factor receptor generate both constitutively active and inactive mutants.

@article{Ishii1997AlanineEO,
  title={Alanine exchanges of polar amino acids in the transmembrane domains of a platelet-activating factor receptor generate both constitutively active and inactive mutants.},
  author={Isao Ishii and Takashi Izumi and Hisao Tsukamoto and Hideaki Umeyama and Michio Ui and Takao Shimizu},
  journal={The Journal of biological chemistry},
  year={1997},
  volume={272 12},
  pages={7846-54}
}
To determine ligand-binding sites of a platelet-activating factor (PAF) receptor, alanine-scanning mutagenesis was carried out. All 23 polar amino acids in the putative 7-transmembrane (TM) domains of a guinea pig PAF receptor were individually replaced with alanine. The ligand-binding properties of mutant receptors were determined after transient expression in COS-7 cells. Mutants in TM II (N58A, D63A), TM III (N100A, T101A, S104A) and TM VII (D289A) displayed higher PAF-binding affinities… CONTINUE READING