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Mutagenesis analysis of the membrane‐proximal ligand binding site of the TGF‐ β receptor type III extracellular domain
Author(s) -
Marie-Claude Pépin,
M Beauchemin,
Catherine Collins,
J. Plamondon,
Maureen D. O’Connor-McCourt
Publication year - 1995
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 - 10.1016/0014-5793(95)01378-4
Subject(s) - binding site , alanine , site directed mutagenesis , mutagenesis , transmembrane domain , asparagine , chemistry , receptor , extracellular , ligand (biochemistry) , biochemistry , amino acid , transmembrane protein , a site , biology , mutation , mutant , gene
There are two TGF‐ β binding subdomains in the extracellular domain of receptor type III (proximal and distal in relation to the transmembrane domain). Here we present an extension of our analysis of the proximal binding site of receptor type III. Due to the original deletion mutagenesis strategy, our proximal binding site contained 19 amino acids from the N‐terminal part of the receptor. By deleting these, we demonstrated that they did not contribute to the binding ability of the proximal binding site. We also produced a soluble, secreted form of the proximal binding site and demonstrated that it was able to bind TGF‐ β . Finally, we analyzed the role of the three asparagine residues (580, 591, 595) that are located in the region of the receptor that is necessary for expression of a functional proximal binding site, and found that mutation of these residues individually to alanine did not affect ligand binding.