Preparation, application and biological characterization of interleukin-1 beta mutant protein biotinylated at a single site.
Selective biotinylation of human interleukin-1 beta was achieved by reaction of an interleukin-1 beta mutant protein containing a surface-accessible cysteine with maleimido-biotin. A defined interleukin-1 beta derivative biotinylated at a single site was obtained. This compound retained full affinity for both the interleukin-1 receptor and streptavidin or avidin; however, its biological activity was significantly reduced in proliferative assays. Biotinylated interleukin-1 beta mutant protein bound to fluorescein-labelled avidin was used in flow cytometric analysis of interleukin-1 receptors. Internalization of the complex between biotinylated interleukin-1 beta mutant protein and streptavidin in a murine thymoma cell line was demonstrated.
Chollet
A
A
Glaxo Institute for Molecular Biology SA., Geneva, Switzerland.
Bonnefoy
J Y
JY
Odermatt
N
N
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D001710:Biotin / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D011971:Receptors, Immunologic / Q000032:analysis* / Q000378:metabolism; D017472:Receptors, Interleukin-1; D019809:Streptavidin
D001426:Bacterial Proteins; D007375:Interleukin-1; D011971:Receptors, Immunologic; D017472:Receptors, Interleukin-1; D001710:Biotin; D019809:Streptavidin