Rational cytokine design for increased lifetime and enhanced potency using pH-activated "histidine switching" Export

@article{sarkar:02, abstract = {We describe a method for the rational design of more effective therapeutic proteins using amino acid substitutions that reduce receptor binding affinity in intracellular endosomal compartments, thereby leading to increased recycling in the ligand-sorting process and consequently resulting in longer half-life in extracellular medium. We demonstrate this approach for granulocyte colony-stimulating factor by using computationally predicted histidine substitutions that switch protonation states between cell-surface and endosomal pH. Molecular modeling of binding electrostatics indicates two different single-histidine mutants that fulfill our design requirements; experimental assays demonstrate that each mutant indeed exhibits an order-of-magnitude increase in medium half-life along with enhanced potency due to increased endocytic recycling.}, author = {Sarkar, Casim A. and Lowenhaupt, Ky and Horan, Thomas and Boone, Thomas C. and Tidor, Bruce and Lauffenburger, Douglas A.}, citeulike-article-id = {3808739}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nbt725}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nbt725}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/12161759}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=12161759}, doi = {10.1038/nbt725}, journal = {Nat Biotech}, keywords = {biology, electrostatics, engineering, protein, systems}, number = {9}, pages = {908--913}, posted-at = {2009-01-08 01:29:49}, priority = {0}, title = {Rational cytokine design for increased lifetime and enhanced potency using pH-activated "histidine switching"}, url = {http://dx.doi.org/10.1038/nbt725}, volume = {20}, year = {2002} }

TY - JOUR ID - sarkar:02 L3 - citeulike-article-id:3808739 N2 - We describe a method for the rational design of more effective therapeutic proteins using amino acid substitutions that reduce receptor binding affinity in intracellular endosomal compartments, thereby leading to increased recycling in the ligand-sorting process and consequently resulting in longer half-life in extracellular medium. We demonstrate this approach for granulocyte colony-stimulating factor by using computationally predicted histidine substitutions that switch protonation states between cell-surface and endosomal pH. Molecular modeling of binding electrostatics indicates two different single-histidine mutants that fulfill our design requirements; experimental assays demonstrate that each mutant indeed exhibits an order-of-magnitude increase in medium half-life along with enhanced potency due to increased endocytic recycling. IS - 9 JF - Nat Biotech EP - 913 TI - Rational cytokine design for increased lifetime and enhanced potency using pH-activated "histidine switching" VL - 20 SP - 908 KW - biology KW - electrostatics KW - engineering KW - protein KW - systems AU - Sarkar, Casim AU - Lowenhaupt, Ky AU - Horan, Thomas AU - Boone, Thomas AU - Tidor, Bruce AU - Lauffenburger, Douglas PY - 2002/// UR - http://dx.doi.org/10.1038/nbt725 ER -