Analysis of eosinophils and myeloid progenitor responses to modified forms of MPIF-2.
Myeloid progenitor inhibitory factor (MPIF)-2 is a beta-chemokine with select and potent activities on eosinophils and myeloid progenitors. In the beta-chemokine family, biological activity is modulated by differential processing of the amino-terminus. Here, for MPIF-2, we describe the biological activities of NH(2)-terminal deletion mutants and compare regions necessary for eosinophil and myeloid progenitor activities. Five MPIF-2 proteins with deletions at the amino-terminus were produced in Escherichia coli and assayed for calcium mobilization, chemotaxis and receptor binding activities on eosinophils, and for their ability to inhibit colony formation of human myeloid bone marrow progenitors. For eosinophils, deletion of the first two amino acids did not markedly alter activity, while subsequent truncations result in a complete loss of activity. One of the MPIF-2 mutants, MPIF-2 (P30-R99) was converted from an agonist to an antagonist of eotaxin, MPIF-2 and MCP-4 functional responses in eosinophil calcium flux and chemotaxis assays. Surprisingly, while displaying a complete loss of agonist activity toward eosinophils, MPIF-2 (P30-R99) retains ability to inhibit human bone marrow myeloid progenitor cell colony formation. In addition, processing at the amino terminus of MPIF-2 in vivo, may result in a chemokine with altered biological activities.
Human Genome Sciences Inc., 9410 Key West Avenue, Rockville, MD 20850, USA.
K J
KJ
Grzegorzewski
X T
XT
Yao
B
B
Kreider
H S
HS
Olsen
T S
TS
Morris
L
L
Zhang
I
I
Sanyal
B
B
Nardelli
D
D
Zukauskas
L
L
Brewer
G W
GW
Bong
Y
Y
Kim
G
G
Garotta
T W
TW
Salcedo
D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D054423:Chemokine CCL24; D019742:Chemokines, CC / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D004804:Eosinophils / Q000378:metabolism*; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D023461:Myeloid Progenitor Cells / Q000378:metabolism*; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D054397:Receptors, CCR3; D019707:Receptors, Chemokine / Q000378:metabolism; D015728:Receptors, HIV / Q000378:metabolism; D017384:Sequence Deletion
C515484:CCL24 protein, human; C515350:CCR3 protein, human; D054423:Chemokine CCL24; D019742:Chemokines, CC; D010446:Peptide Fragments; D054397:Receptors, CCR3; D019707:Receptors, Chemokine; D015728:Receptors, HIV; D002118:Calcium