High-resolution epitope mapping of hGH-receptor interactions by alanine-scanning mutagenesis

@article{citeulike:1223150, abstract = {A strategy, called alanine-scanning mutagenesis, was used to identify specific side chains in human growth hormone (hGH) that strongly modulate binding to the hGH receptor cloned from human liver. Single alanine mutations (62 in total) were introduced at every residue contained within the three discontinuous segments of hGH (residues 2 to 19, 54 to 74, and 167 to 191) that have been implicated in receptor recognition. The alanine scan revealed a cluster of a dozen large side chains that when mutated to alanine each showed more than a four times lower binding affinity to the hGH receptor. Many of these residues that promote binding to the hGH receptor are altered in homologs of hGH (such as placental lactogens and prolactins) that do not bind tightly to the hGH receptor. The overall folding of these mutant proteins was indistinguishable from that of the wild-type hGH, as determined by strong cross-reactivities with seven different conformationally sensitive monoclonal antibodies. The alanine scan also identified at least one side chain, Glu174, that hindered binding because when it was mutated to alanine the receptor affinity increased by more than a factor of four. 10.1126/science.2471267}, author = {Cunningham, Bc and Wells, Ja }, citeulike-article-id = {1223150}, doi = {http://dx.doi.org/10.1126/science.2471267}, journal = {Science}, keywords = {combinatorial, function, mutagenesis}, month = {June}, number = {4908}, pages = {1081--1085}, posted-at = {2007-04-13 00:34:26}, priority = {2}, title = {High-resolution epitope mapping of hGH-receptor interactions by alanine-scanning mutagenesis}, url = {http://dx.doi.org/10.1126/science.2471267}, volume = {244}, year = {1989} }

TY - JOUR ID - citeulike:1223150 L3 - citeulike-article-id:1223150 TI - High-resolution epitope mapping of hGH-receptor interactions by alanine-scanning mutagenesis JF - Science VL - 244 IS - 4908 SP - 1081 EP - 1085 N2 - A strategy, called alanine-scanning mutagenesis, was used to identify specific side chains in human growth hormone (hGH) that strongly modulate binding to the hGH receptor cloned from human liver. Single alanine mutations (62 in total) were introduced at every residue contained within the three discontinuous segments of hGH (residues 2 to 19, 54 to 74, and 167 to 191) that have been implicated in receptor recognition. The alanine scan revealed a cluster of a dozen large side chains that when mutated to alanine each showed more than a four times lower binding affinity to the hGH receptor. Many of these residues that promote binding to the hGH receptor are altered in homologs of hGH (such as placental lactogens and prolactins) that do not bind tightly to the hGH receptor. The overall folding of these mutant proteins was indistinguishable from that of the wild-type hGH, as determined by strong cross-reactivities with seven different conformationally sensitive monoclonal antibodies. The alanine scan also identified at least one side chain, Glu174, that hindered binding because when it was mutated to alanine the receptor affinity increased by more than a factor of four. 10.1126/science.2471267 KW - combinatorial KW - function KW - mutagenesis AU - Cunningham, Bc AU - Wells, Ja PY - 1989/06/02/ UR - http://dx.doi.org/10.1126/science.2471267 ER -