Journal home > Archive > Table of Contents > Research >  Article > Abstract

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 Methods of Modeling Adaptation in Populations Deadline: Dec 30 2009 Reward: $15,000 USD The analysis of adaptation with a population is a frequently encountered computational modeling scen... Single-cell Analysis Platform Deadline: Dec 02 2009 Reward: $5,000 USD This Challenge is looking for novel approaches to analyzing changes at a single-cell level. This is... More Challenges Powered by: nature jobs Chief Scientific Manager - Medicinal Chemistry Syngene International Bangalore, Karnataka 560099 India Senior Research Fellow - Atlantic Ocean Circulation and Climate University of Southampton Southampton / Hampshire United Kingdom More science jobs Post a job for free Figures & Tables See also: Research News by Robinson Export citation Search buyers guide:   ADVERTISEMENT

ISSN: 1087-0156 EISSN: 1546-1696 Journal home | Advance online publication | Current issue | Archive | Press releases | Supplements | Focuses | Conferences | For authors | Online submission | Permissions | For referees | Free online issue | About the journal | Contact the journal | Subscribe | Advertising | work@npg | naturereprints | About this site | For librarians

©2002 Nature Publishing Group | Privacy policy