title: Regulation and function of the protein inhibitor of nitric oxide synthase (PIN)/dynein light chain 8 (LC8) in a human mast cell line.
abstract: The protein inhibitor of nitric oxide synthase (PIN) was independently identified as an inhibitor of nitric oxide (NO) produced by neuronal nitric oxide synthase (nNOS), and as a member of the cellular dynein light chain family, dynein light chain 8 (LC8), responsible for intracellular protein trafficking. Mast cells (MC) are involved in several homeostatic and pathological processes and can be regulated by NO. This study describes the expression of PIN/LC8 in the human MC line HMC-1. We also studied if PIN/LC8 binds nNOS, and what role this might have in leukotriene (LT) production. We found that PIN/LC8 mRNA and protein was expressed in HMC-1. Using a GST-PIN construct, we showed PIN binds to nNOS, but not endothelial (e)NOS in HMC-1; in our studies HMC-1 did not express inducible (i)NOS. Intracellular delivery of anti-PIN/LC8 antibody enhanced ionophore (A23187)-induced LT production through an unknown mechanism. Thus we established for the first time expression of PIN/LC8 in human MC, its ability to bind nNOS, and the effect that blocking it has on LT production in a human MC lines.
authors.affiliation: Glaxo-Heritage Asthma Research Laboratory, Pulmonary Research Group, Department of Medicine, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Scott D
authors.initials: SD
authors.lastname: McCauley
authors.forename: Mark
authors.initials: M
authors.lastname: Gilchrist
authors.forename: A Dean
authors.initials: AD
authors.lastname: Befus
mesh_terms: D000818:Animals; D015153:Blotting, Western; D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D056445:Cytoplasmic Dyneins; D004398:Dyneins / Q000096:biosynthesis*; D004926:Escherichia coli / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D015289:Leukotrienes / Q000096:biosynthesis; D008407:Mast Cells / Q000502:physiology*; D051379:Mice; D052248:Nitric Oxide Synthase Type I / Q000037:antagonists & inhibitors*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011863:Radioimmunoassay; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D007074:Immunoglobulin G; D015289:Leukotrienes; D012333:RNA, Messenger; D000001:Calcimycin; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C427758:DYNLL1 protein, human; D056445:Cytoplasmic Dyneins; D004398:Dyneins; D005978:Glutathione