Electrospray ionization mass spectrometric analyses of phospholipids from rat and human pancreatic islets and subcellular membranes: comparison to other tissues and implications for membrane fusion in insulin exocytosis.
Glucose-induced insulin secretion from pancreatic islets involves hydrolysis of arachidonic acid from phospholipids as an intermediary event. Accumulation of nonesterified arachidonate in islet membranes may influence both ion fluxes that trigger insulin secretion and fusion of secretory granule and plasma membranes. Recent findings indicate that plasmenylethanolamine species may also participate in fusion of such membranes, but high-performance liquid chromatographic (HPLC) and gas chromatographic/mass spectrometric (GC/MS) analyses of islet secretory granule phospholipids suggested that they contain little plasmenylethanolamine. Here, electrospray ionization mass spectrometry (ESI/MS) of intact phospholipid molecules is used to demonstrate that the most prominent components of all major glycerophospholipid headgroup classes in islets are arachidonate-containing species. Such species contribute the majority of the ESI/MS negative ion current from rat and human islet glycerophosphoethanolamine (GPE), and the fraction of GPE negative ion current contributed by plasmenylethanolamine species in rat islets is higher than that for rat liver or heart and similar to that for brain. The most prominent sn-2 substituent of plasmenylethanolamine species in brain is docosahexaenoate and in islets is arachidonate. Arachidonate-containing plasmenylethanolamine species are also prominent components of GPE from islet secretory granules and plasma membranes. Fusion of islet secretory granule and plasma membranes is demonstrated to be catalyzed by cytosolic components from insulinoma cells and rat brain with chromatographic similarities to a rabbit brain factor that specifically catalyzes fusion of plasmenylethanolamine-containing membranes.
Ramanadham
S
S
Mass Spectrometry Resource, Division of Diabetes, Endocrinology, and Metabolism, Department of Medicine, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, Missouri 63110, USA.
Hsu
F F
FF
Bohrer
A
A
Nowatzke
W
W
Ma
Z
Z
Turk
J
J
D000818:Animals; D016718:Arachidonic Acid / Q000737:chemistry; D001921:Brain / Q000378:metabolism; D004281:Docosahexaenoic Acids / Q000737:chemistry; D005089:Exocytosis; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007425:Intracellular Membranes / Q000737:chemistry / Q000378:metabolism; D007515:Islets of Langerhans / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D008561:Membrane Fusion; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D007328:Insulin; D008563:Membrane Lipids; D010714:Phosphatidylethanolamines; D010743:Phospholipids; C002449:glycerophosphoethanolamine; D004281:Docosahexaenoic Acids; D016718:Arachidonic Acid