RT Journal Article
SR Electronic
T1 Rational Design of a Bifunctional AND-Gate Ligand to Modulate Cell-Cell Interactions
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 711549
DO 10.1101/711549
A1 Jungmin Lee
A1 Andyna Vernet
A1 Katherine Redfield
A1 Shulin Lu
A1 Ionita C. Ghiran
A1 Jeffrey C. Way
A1 Pamela A. Silver
YR 2019
UL http://biorxiv.org/content/early/2019/07/23/711549.abstract
AB Protein “AND-gate” systems, in which a ligand acts only on cells with two different receptors, direct signaling activity to a particular cell type and avoid action on other cells. In a bifunctional AND-Gate protein, the molecular geometry of the protein domains is crucial. Here we constructed a tissue-targeted erythropoietin (EPO) that stimulates red blood cell (RBC) production without triggering thrombosis. EPO was directed to RBC precursors and mature RBCs by fusion to an anti-glycophorin A antibody V region. Many such constructs activated EPO receptors in vitro and stimulated RBC and not platelet production in mice but nonetheless enhanced thrombosis in mice and caused adhesion between RBCs and EPO receptor-bearing cells. Based on a protein-structural model of the RBC surface, we rationally designed an anti-glycophorin/EPO fusion that does not induce cell adhesion in vitro or enhance thrombosis in vivo. Thus, meso-scale geometry can inform design of synthetic-biological systems.