A peptide with unique receptor specificity: stimulation of phosphoinositide hydrolysis and induction of superoxide generation in human neutrophils.
Previously, we identified peptides that stimulate phosphoinositide hydrolysis in several leukocyte cell lines from mixtures of random hexapeptide sequences. Moreover, the peptides activate phospholipase C via a pertussis toxin-sensitive G protein-coupled receptor. We now investigate the structure-activity relationship of the peptides with the goal of improving the activity of the peptides, as well as the biologic function of the peptides. Substitution of the L-methionine at the C terminus of peptides with D-methionine markedly increased the effectiveness of the peptides. The half-maximal effective concentrations of MKYMPm-NH2 and WKYMVm-NH2 for stimulation of phosphoinositide hydrolysis in U266 cells were 30 and 0.5 nM, respectively. By BIAcore analysis we confirmed the existence of a receptor for WKYMVm-NH2. Furthermore, the intracellular calcium concentration increase induced by WKYMVm-NH2 was not inhibited by several chemoattractants (FMLP, IL-8, platelet-activating factor, C5a, granulocyte-macrophage CSF, and granulocyte CSF) suggests that WKYMVm-NH2 has a unique cell surface receptor on leukocytes. WKYMVm-NH2 stimulated the phosphoinositide hydrolysis in U937, HL60, and U266 cells, as well as in human neutrophils. Moreover, WKYMVm-NH2 is more effective than FMLP in the production of superoxide in human neutrophils. The data suggest that WKYMVm-NH2 may have the ability to activate the microbicidal functions of human neutrophils.
Seo
J K
JK
Department of Life Science, Pohang University of Science and Technology, Hyojadong, Korea.
Choi
S Y
SY
Kim
Y
Y
Baek
S H
SH
Kim
K T
KT
Chae
C B
CB
Lambeth
J D
JD
Suh
P G
PG
Ryu
S H
SH
D000596:Amino Acids / Q000737:chemistry; D015374:Biosensing Techniques; D002118:Calcium / Q000378:metabolism; D005998:Glycine / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000187:drug effects; D006801:Humans; D006868:Hydrolysis; D007424:Intracellular Fluid / Q000378:metabolism; D016393:Lymphoma, B-Cell; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D009842:Oligopeptides / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology*; D010716:Phosphatidylinositols / Q000378:metabolism*; D018000:Receptors, Peptide / Q000378:metabolism / Q000502:physiology*; D017384:Sequence Deletion; D013237:Stereoisomerism; D013481:Superoxides / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D009842:Oligopeptides; D010716:Phosphatidylinositols; D018000:Receptors, Peptide; D013481:Superoxides; D002118:Calcium; D005998:Glycine