Structure-function analysis of FLT3 ligand-FLT3 receptor interactions using a rapid functional screen.

The Journal of Biological Chemistry
T J GraddisJ T McGrew

Abstract

FLT3 ligand (FLT3L) stimulates primitive hematopoietic cells by binding to and activating the FLT3 receptor (FLT3R). We carried out a structure-activity study of human FLT3L in order to define the residues involved in receptor binding. We developed a rapid method to screen randomly mutagenized FLT3L using a FLT3R-Fc fusion protein to probe the relative binding activities of mutated ligand. Approximately 60,000 potential mutants were screened, and the DNA from 59 clones was sequenced. Thirty-one single amino acid substitutions at 24 positions of FLT3L either enhanced or reduced activity in receptor binding and cell proliferation assays. Eleven representative proteins were purified and analyzed for receptor affinity, specific activity, and physical properties. Receptor affinity and bioactivity were highly correlated. FLT3L affinity for receptor improved when four individual mutations that enhance FLT3L receptor affinity were combined in a single molecule. A model of FLT3L three-dimensional structure was generated based on sequence alignment and x-ray structure of macrophage colony-stimulating factor. Most residues implicated in receptor binding are widely dispersed in the primary structure of FLT3L, yet they localize to a surface...Continue Reading

References

Feb 1, 1992·Yeast·D MuhlradR Parker
Jan 1, 1991·Methods in Enzymology·F Sherman
Aug 24, 1990·Cell·R D Klausner, R Sitia
Jan 18, 1994·Proceedings of the National Academy of Sciences of the United States of America·D SmallA M Gewirtz
Mar 15, 1994·Structure·D A RozwarskiP A Karplus
Feb 1, 1993·Protein Science : a Publication of the Protein Society·S SrinivasanS Sudarsanam
Aug 1, 1995·International Journal of Hematology·S D Lyman
Mar 7, 1997·The Journal of Biological Chemistry·M A LemmonJ Schlessinger
Dec 1, 1996·Nature Biotechnology·C C ReddyD A Lauffenburger

❮ Previous
Next ❯

Citations

Feb 3, 2009·The Protein Journal·Kenneth VerstraeteSavvas N Savvides
Oct 19, 2012·Nature Reviews. Cancer·Kenneth Verstraete, Savvas N Savvides
Oct 27, 1999·Proceedings of the National Academy of Sciences of the United States of America·A CarfíD C Wiley
Feb 14, 2003·Clinical Immunology : the Official Journal of the Clinical Immunology Society·Edgar G Engleman, Lawrence Fong
Dec 18, 2009·Developmental and Comparative Immunology·Laurence Guzylack-PiriouArtur Summerfield
Oct 7, 2011·Cytokine & Growth Factor Reviews·Anna TarasovaDavid Winkler
May 9, 2019·Physiological Reviews·Julhash U Kazi, Lars Rönnstrand
Feb 21, 2003·The Journal of Immunology : Official Journal of the American Association of Immunologists·Gangadhara SailajaAbdul M Jabbar
Oct 20, 2007·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Francesca OrlandiPolly D Gregor
Apr 9, 2021·Acta Crystallographica. Section F, Structural Biology Communications·Erwin PannecouckeSavvas N Savvides
Jul 6, 2000·Nature Structural Biology·C Wiesmann, A M de Vos
Jul 6, 2000·Nature Structural Biology·S N SavvidesP Andrew Karplus

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.

Cell Signaling by Tyrosine Kinases

Receptor tyrosine kinases (RTKs) are the high-affinity cell surface receptors for many polypeptide growth factors, cytokines, and hormones. RTKs have been shown not only to be key regulators of normal cellular processes but also to have a critical role in the development and progression of many types of cancer. Discover the latest research on cell signaling and RTK here.