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

Authors :
Sarkar CA
Lowenhaupt K
Horan T
Boone TC
Tidor B
Lauffenburger DA
Source :
Nature biotechnology [Nat Biotechnol] 2002 Sep; Vol. 20 (9), pp. 908-13. Date of Electronic Publication: 2002 Aug 05.
Publication Year :
2002

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.

Details

Language :
English
ISSN :
1087-0156
Volume :
20
Issue :
9
Database :
MEDLINE
Journal :
Nature biotechnology
Publication Type :
Academic Journal
Accession number :
12161759
Full Text :
https://doi.org/10.1038/nbt725