Role of iduronate-2-sulfatase in glucose-stimulated insulin secretion by activation of exocytosis.
Iduronate-2-sulfatase (IDS) is a lysosomal enzyme expressed in pancreatic islets responsible for the degradation of proteoglycans such as perlecan and dermatan sulfate. Previous findings of our group demonstrated the involvement of IDS in the normal pathway of lysosomal degradation of secretory peptides, suggesting a role of this enzyme in beta-cell secretory functionality. The present study was undertaken to characterize the effect of IDS overexpression on insulin release. INS1E cells were transiently transfected with a construct encoding human IDS (hIDS). hIDS overexpression was associated with a gain of function detected by a reduction in heparan sulfate content. hIDS potentiated the glucose-stimulated insulin secretory response compared with controls (61%) with no changes in insulin mRNA levels or insulin peptide content. Results on quantification of the exocytotic process showed a significant increase in hIDS-transfected cells compared with controls. Furthermore, ultramorphological analysis demonstrated an increase in the number of granules in the immediate vicinity of the plasma membrane in hIDS-transfected cells and a decrease in total vesicles per square micrometer. hIDS overexpression induced phosphorylation of protein kinase C (PKC) alpha and its newly myristoylated alanine-rich C kinase substrate, MARCKS. We conclude that IDS has a role in glucose-stimulated insulin secretion via a mechanism that involves the activation of exocytosis through phosphorylation of PKCalpha and MARCKS.
Piquer
S
S
Endocrinology and Diabetes Unit, Laboratory of Diabetes and Obesity, Institut d' Investigacions Biomediques August Pi iSunyer-Fundació Clínic, Hospital Clínic, Barcelona, Spain.
Casas
S
S
Quesada
I
I
Nadal
A
A
Julià
M
M
Novials
A
A
Gomis
R
R
D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D005089:Exocytosis / Q000187:drug effects* / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D007066:Iduronate Sulfatase / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism / Q000502:physiology; D007515:Islets of Langerhans / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D051571:Protein Kinase C-alpha / Q000378:metabolism / Q000502:physiology; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics / Q000502:physiology
D007328:Insulin; D047908:Intracellular Signaling Peptides and Proteins; C000619710:MARCKS protein, human; D008565:Membrane Proteins; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D051571:Protein Kinase C-alpha; D007066:Iduronate Sulfatase; D005947:Glucose