IL-8 derivatives with a reduced potential to form homodimers are fully active in vitro and in vivo.
Interleukin 8 (IL-8) is a member of the CXC subfamily of chemokines which attracts and activates preferentially neutrophilic granulocytes. At nanomolar concentrations monomeric and dimeric forms of the molecule are in equilibrium, with the monomer being the prevalent form. Five amino acids from position 23 to 29 of the 72-amino acid IL-8 sequence form the dimer interface, with Leu25 and Val27 being highly conserved among the CXC chemokines. To investigate the contribution of these amino acids to the dimerization of IL-8, we produced in escherichia coli IL-8 derivatives with phenylalanine substitutions at position 25 or 27, or both 25 and 27. All three recombinant proteins were characterized by a significantly impaired potential to form dimers in solution as seen in chemical crosslinking experiments. IL-8 Val27 also could not be crosslinked as a dimer on its receptors. Receptor affinities and in vitro chemotactic activities, however, were not significantly different between wild-type and IL-8 with single mutations. The dimerization deficient IL-8 analogue had also full inflammatory activity in vivo. Thus, the monomer is the biologically active form of IL-8.
Horcher
M
M
Sandoz Forschungsinstitut GmbH, Vienna, Austria.
Rot
A
A
Aschauer
H
H
Besemer
J
J
D015703:Antigens, CD / Q000378:metabolism; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000378:metabolism; D054407:Chemokine CCL4; D019281:Dimerization; D006801:Humans; D016209:Interleukin-8 / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D019402:Macrophage Inflammatory Proteins / Q000378:metabolism; D016296:Mutagenesis; D018123:Receptors, Interleukin / Q000378:metabolism; D023062:Receptors, Interleukin-8A
D015703:Antigens, CD; D018932:Chemokine CCL2; D054407:Chemokine CCL4; D016209:Interleukin-8; D019402:Macrophage Inflammatory Proteins; D018123:Receptors, Interleukin; D023062:Receptors, Interleukin-8A