vasoactive-intestinal-peptide and Hemolysis

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a naturally occurring cationic peptide with potent immunosuppressant and cytoprotective activities. We now show that full length PACAP38 and to a lesser extent, the truncated form PACAP27, and the closely related vasoactive intestinal peptide (VIP) and secretin had antimicrobial activity against the Gram-negative bacteria Escherichia coli in the radial diffusion assay. PACAP38 was more potent than either the bovine neutrophil antimicrobial peptide indolicidin or the synthetic antimicrobial peptide ARVA against E. coli. PACAP38 also had activity against the Gram-positive bacteria Staphylococcus aureus in the same assay with comparable potency to indolicidin and ARVA. In the more stringent broth dilution assay, PACAP38 had moderate sterilizing activity against E. coli, and potent sterilizing activity against the Gram-negative bacteria Pseudomonas aeruginosa. PACAP27, VIP and secretin were much less active than PACAP38 in this assay. PACAP38 also had some activity against the Gram-positive bacteria Bacillus cereus in the broth dilution assay. Many exopeptidase-resistant analogs of PACAP38, including both receptor agonists and antagonists, had antimicrobial activities equal to, or better than PACAP38, in both assays. PACAP38 made the membranes of E. coli permeable to SYTOX Green, suggesting a classical membrane lytic mechanism. These data suggest that analogs of PACPAP38 with a wide range of useful biological activities can be made by judicious substitutions in the sequence.

Topics: Anti-Infective Agents; Cell Membrane; Chromatography, High Pressure Liquid; Escherichia coli; Hemolysis; Pituitary Adenylate Cyclase-Activating Polypeptide; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Structure-Activity Relationship; Vasoactive Intestinal Peptide

1 Vasoactive intestinal polypeptide (VIP, 0.01- MicroM) produced dose-related relaxations of the mouse anococcygeus muscle. 2 Following incubation with indomethacin (2.8 microM 1 h) adenosine 5'-triphosphate (ATP, 0.5-10 mM) produced dose-related relaxations of the mouse anococcygeus. 3 Haemolysed blood reduced inhibitory responses of the mouse anococcygeus to field stimulation but had no effect on relaxations to VIP or ATP. 4 Apamin (0.5 microM) had no effect on the relaxation of mouse anococcygeus to field stimulation, VIP, or ATP. 5 2-2'-Pyridylisatogen tosylate (PIT, 50 microM) itself reduced muscle tone but it did not abolish inhibitory responses to field stimulation, VIP, or ATP. 6 During prolonged inhibitory nerve stimulation the relaxation of the mouse anococcygeus in response to VIP was reduced greatly while that to ATP was unaffected. 7 Bundles of VIP-immunoreactive sites were detected in sections of the mouse anococcygeus treated by the peroxidase-antiperoxidase (PAP) immunocytochemical technique. 8 The results suggest that the mechanisms underlying non-adrenergic, non-cholinergic inhibitory transmission in the mouse anococcygeus are similar to those in the bovine retractor penis and unlike those in the guinea-pig taenia caeci. 9 The possibility that VIP or ATP might be involved in inhibitory neurotransmission in the mouse anococcygeus is discussed.

Topics: Adenosine Triphosphate; Animals; Apamin; Blood Physiological Phenomena; Electric Stimulation; Gastrointestinal Hormones; Hemolysis; In Vitro Techniques; Isatin; Male; Mice; Muscle Contraction; Muscle Relaxation; Radioimmunoassay; Vasoactive Intestinal Peptide