Effects of tripeptides derived from milk proteins on polymorphonuclear oxidative and phosphoinositide metabolisms.
The tripeptide GLF (glycyl-leucyl-phenylalanine) was isolated from human milk proteins. This peptide increased phagocytosis by human and murine macrophages and protected mice against Klebsiella pneumoniae infection. Specific binding sites on human polymorphonuclear leukocytes (PMNs) have been demonstrated recently. The aim of the present research was to study the action of this peptide on rat and human PMN oxidative burst and to investigate the consequences of cell stimulation on polyphosphoinositide hydrolysis. A biphasic stimulating concentration-dependent effect of GLF on PMN chemiluminescence and superoxide anion generation was demonstrated. One of the peaks of the oxidative response occurred around 10(-9) M, which correlates with the Kd of high affinity receptors of GLF. The other maximum, around 10(-4) M, might be due to the hydrophobic nature of the tripeptide. O2- generation mimicked the phorbol myristate acetate response: after a lag period of 2-5 min, O2- release gradually increased for 10-15 min until a plateau was reached. Furthermore, GLF enhanced phosphoinositide breakdown with maximal IP3 production at 10(-7) M. Various analogs of GLF were synthesized in order to define the relative importance of the different amino acids and their position in the tripeptide molecule: glycyl-phenylalanine-leucine was devoid of biological properties but enhanced the activity of GLF on the metabolic burst at high concentrations; peptides leucyl-leucyl-phenylalanine and leucyl-leucyl-tyrosine, which displaced GLF from its specific membrane receptors, exerted stimulating effects on PMN oxidative and phosphoinositide metabolisms. It is quite conceivable that these short peptides, which may be generated in the newborn during digestion and which are able to stimulate phagocytic cells, are implicated in the defense of the neonate immature organism against infection.
Migliore-Samour
D
D
Laboratoire des Protéines, CNRS URA 1188, Université de Paris V, France.
Roch-Arveiller
M
M
Tissot
M
M
Jazziri
M
M
Keddad
K
K
Giroud
J P
JP
Jollès
P
P
D000818:Animals; D006801:Humans; D008163:Luminescent Measurements; D008297:Male; D008894:Milk Proteins / Q000737:chemistry / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects* / Q000502:physiology; D009842:Oligopeptides / Q000302:isolation & purification / Q000494:pharmacology*; D010587:Phagocytosis / Q000187:drug effects; D010716:Phosphatidylinositols / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D016897:Respiratory Burst / Q000187:drug effects; D013481:Superoxides / Q000378:metabolism*; D015054:Zymosan / Q000494:pharmacology
D008894:Milk Proteins; D009842:Oligopeptides; D010716:Phosphatidylinositols; C062537:glycyl-leucyl-phenylalanine; D013481:Superoxides; D015054:Zymosan