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Journal home Advance online publication Current issue Archive Press releases Supplements Focuses Conferences Guide to authors Online submission Permissions For referees Free online issue Contact the journal Subscribe Advertising work@npg naturereprints About this site For librarians   NPG Resources Bioentrepreneur Nature Reviews Drug Discovery Nature Nature Medicine Nature Genetics Nature Reviews Genetics Nature Methods Nature Chemical Biology news@nature.com Clinical Pharmacology & Therapeutics Nature Conferences NPG Subject areas Biotechnology Cancer Chemistry Clinical Medicine Dentistry Development Drug Discovery Earth Sciences Evolution & Ecology Genetics Immunology Materials Science Medical Research Microbiology Molecular Cell Biology Neuroscience Pharmacology Physics Browse all publications Article Nature Biotechnology  20, 908 - 913 (2002) Published online: 5 August 2002; | doi:10.1038/nbt725 Rational cytokine design for increased lifetime and enhanced potency using pH-activated "histidine switching" Casim A. Sarkar1, 2, Ky Lowenhaupt3, Thomas Horan6, Thomas C. Boone6, Bruce Tidor4, 5 & Douglas A. Lauffenburger1, 2, 3, 4 1  Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139-4307. 2  Biotechnology Process Engineering Center, Massachusetts Institute of Technology, Cambridge, MA 02139-4307. 3  Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139-4307. 4  Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, MA 02139-4307. 5  Department of Electrical Engineering & Computer Science, Massachusetts Institute of Technology, Cambridge, MA 02139-4307. 6  Department of Process Science, Amgen, Inc., Amgen Center, Thousand Oaks, CA 91320-1789. Correspondence should be addressed to Douglas A. Lauffenburger lauffen@mit.edu or Bruce Tidor tidor@mit.eduWe 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. MORE ARTICLES LIKE THIS These links to content published by NPG are automatically generated NEWS AND VIEWS Reduce, reuse, and recycle Nature Biotechnology Research News (01 Sep 2002) RESEARCH Hrs sorts ubiquitinated proteins into clathrin-coated microdomains of early endosomes Nature Cell Biology Brief Communication (01 May 2002)  Top Abstract Previous | Next Table of contents Full text Download PDF Send to a friend Save this link Open Innovation Challenges Mitigating Zinc Corrosion Deadline: Aug 23 2009 Reward: $20,000 USD The Seeker is looking for novel methods to mitigate zinc corrosion/gassing in alkaline media. This ... Protect Enzyme from In Planta Degradation Deadline: Jul 15 2009 Reward: $20,000 USD A proposal for stable expression of an enzyme in corn seed is desired. More Challenges Powered by: nature jobs Professorship Friedrich-Alexander-Universitat Erlangen-Nurnberg Erlangen 91054 Germany Research Scientist - Phd - Genomics - Primary Cell Lines KSR Richmond, VA, USA More science jobs Post a job for free Figures & Tables See also: Research News by Robinson Export citation Search buyers guide:   ADVERTISEMENT

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