Identification of a chemotactic domain of the pro-inflammatory S100 protein CP-10.
We previously reported the purification and partial amino acid sequence of a novel murine cytokine designated CP-10, which has chemotactic activity for murine polymorphonuclear cells (PMN) and macrophages. The complete cDNA encoding an 88-amino acid polypeptide has been isolated and the sequence is presented here. Transient transfection of CP-10 cDNA into CV-1 cells confirmed the chemotactic activity of rCP-10 for murine PMN. CP-10 has sequence homology with members of the S100 family of Ca(2+)-binding proteins with pronounced amino acid sequence similarities within the putative N- and C-terminal Ca(2+)-binding sites, but differences within their connecting hinge and C-terminal regions. We have confirmed the hypothesis of Kligman and Hilt that functional specificity of individual members of the S100 protein family may reside in the hinge region. A synthetic peptide corresponding to the hinge region of CP-10 (CP-10(42-55) was compared with native CP-10 in chemotaxis and skin test assays. Native CP-10 had potent activity for phagocytic cells, but not lymphocytes, in vitro (optimal activity, 10(-11) to 10(-13) M) and elicited a sustained recruitment of neutrophils and mononuclear cells over 24 h in vivo. The hinge-region peptide had strong chemotactic activity for murine phagocytic cells (optimal activity, 10(-10) - 10(-11) M) but elicited only a transient infiltration of neutrophils over 4 to 8 h after intradermal injection. Results indicate that although the hinge region contributes significantly to the functional specificity of the S100 protein CP-10, sustained cellular recruitment typical of a delayed type hypersensitivity response is apparently dependent on the structural integrity of the protein.
Lackmann
M
M
Heart Research Institute, Camperdown, NSW, Australia.
Rajasekariah
P
P
Iismaa
S E
SE
Jones
G
G
Cornish
C J
CJ
Hu
S
S
Simpson
R J
RJ
Moritz
R L
RL
Geczy
C L
CL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D054357:Chemokine CXCL10; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D016207:Cytokines / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D004247:DNA / Q000302:isolation & purification; D051379:Mice; D008805:Mice, Inbred A; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley; D009418:S100 Proteins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D013329:Structure-Activity Relationship
D054357:Chemokine CXCL10; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D016207:Cytokines; D009418:S100 Proteins; D004247:DNA