Regulation and function of the protein inhibitor of nitric oxide synthase (PIN)/dynein light chain 8 (LC8) in a human mast cell line.
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.
McCauley
Scott D
SD
Glaxo-Heritage Asthma Research Laboratory, Pulmonary Research Group, Department of Medicine, University of Alberta, Edmonton, Alberta, Canada.
Gilchrist
Mark
M
Befus
A Dean
AD
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
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