Granule localization of glutaminase in human neutrophils and the consequence of glutamine utilization for neutrophil activity.
The provision of glutamine in vivo has been observed to reduce to normal levels the neutrophilia observed after exhaustive exercise and to decrease the neutrophil chemoattractant, interleukin-8. Thus, the role for glutamine in the regulation of inflammatory mediators of human neutrophil activation was investigated. The study sought to establish whether glutamine supplementation in vitro affects neutrophil function at rest and whether glutaminase, the major enzyme that metabolizes glutamine, is present in human polymorphonuclear neutrophils (PMN). During in vitro studies, the addition of 2 mm glutamine increased the respiratory burst of human PMN stimulated with both phorbol myristate acetate (PMA) and formyl-methionyl-leucyl-phenylalanine. These observations were made using a highly sensitive, real time chemiluminescent probe, Pholasin. Glutamine alone did not stimulate the release of reactive oxygen species. In a novel finding using glutaminase-specific antibodies in combination with flow cytometry and confocal microscopy, glutaminase was shown to be present on the surface of human PMN. Subcellular fractionation revealed that the enzyme was enriched in the secondary granules and could be released into cell culture medium upon stimulation with PMA. In conclusion, human PMN appeared to utilize glutamine and possess the appropriate glutaminase enzyme for metabolizing glutamine. This may depress some pro-inflammatory factors that occur during prolonged, exhaustive exercise.
Castell
Linda
L
Department of Biochemistry, University of Oxford, United Kingdom. lindy.castell@nda.ox.ac.uk
Vance
Caroline
C
Abbott
Rachel
R
Marquez
Javier
J
Eggleton
Paul
P
D003594:Cytoplasmic Granules / Q000201:enzymology*; D005972:Glutaminase / Q000378:metabolism*; D005973:Glutamine / Q000493:pharmacokinetics*; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D018375:Neutrophil Activation / Q000187:drug effects / Q000502:physiology*; D009504:Neutrophils / Q000201:enzymology* / Q000378:metabolism; D010084:Oxidation-Reduction; D016897:Respiratory Burst / Q000187:drug effects / Q000502:physiology
D005973:Glutamine; D005972:Glutaminase