RRC ID 41565
Author Hayashi M, Tomita M, Yoshizato K.
Title Interleukin-2-collagen chimeric protein which liberates interleukin-2 upon collagenolysis.
Journal Protein Eng
Abstract Interleukin-2 (IL-2) is a potent activator of cellular immunity and has been utilized as an immunotherapeutic agent. We stably immobilized human IL-2 to collagen by covalently binding it to the N-terminus of human type III collagen (3A1) as IL2-3A1 chimeric protein using recombinant technology. The present study was aimed at liberating IL-2 from the immobilized chimeric protein by treating the chimera with bacterial collagenase. These IL2-3A1 chimeras were synthesized in insect cells which had been infected with baculovirus vectors carrying IL2-3A1 cDNA. The IL2-3A1 protein produced was shown to be in a pepsin-resistant triple helical structure and exhibited IL-2 activity to a similar extent as IL-2 itself. IL2-3A1 could be immobilized on the surface of plastic dishes by incubating it in the dishes. The IL-2 region of the immobilized IL2-3A1 was liberated to culture media by collagenase treatment and this freed IL-2 stimulated the growth of lined T cells. Thus, IL2-3A1 chimeric protein could be utilized as an IL-2 deliverer whose T cell mitogenic activity can be liberated by a collagenolytic environment.
Volume 15(5)
Pages 429-36
Published 2002-5-1
DOI 10.1093/protein/15.5.429
PMID 12034863
MeSH Animals Cell Line Cells, Immobilized / metabolism Chromatography, Affinity Collagen / genetics* Collagen / metabolism Collagenases / metabolism Interleukin-2 / genetics* Interleukin-2 / metabolism Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Spodoptera
Times Cited 8
Human and Animal Cells Sf9(RCB0563)