Role for malic enzyme, pyruvate carboxylation, and mitochondrial malate import in glucose-stimulated insulin secretion.
Pyruvate cycling has been implicated in glucose-stimulated insulin secretion (GSIS) from pancreatic beta-cells. The operation of some pyruvate cycling pathways is proposed to necessitate malate export from the mitochondria and NADP(+)-dependent decarboxylation of malate to pyruvate by cytosolic malic enzyme (ME1). Evidence in favor of and against a role of ME1 in GSIS has been presented by others using small interfering RNA-mediated suppression of ME1. ME1 was also proposed to account for methyl succinate-stimulated insulin secretion (MSSIS), which has been hypothesized to occur via succinate entry into the mitochondria in exchange for malate and subsequent malate conversion to pyruvate. In contrast to rat, mouse beta-cells lack ME1 activity, which was suggested to explain their lack of MSSIS. However, this hypothesis was not tested. In this report, we demonstrate that although adenoviral-mediated overexpression of ME1 greatly augments GSIS in rat insulinoma INS-1 832/13 cells, it does not restore MSSIS, nor does it significantly affect GSIS in mouse islets. The increase in GSIS following ME1 overexpression in INS-1 832/13 cells did not alter the ATP-to-ADP ratio but was accompanied by increases in malate and citrate levels. Increased malate and citrate levels were also observed after INS-1 832/13 cells were treated with the malate-permeable analog dimethyl malate. These data suggest that although ME1 overexpression augments anaplerosis and GSIS in INS-1 832/13 cells, it is not likely involved in MSSIS and GSIS in pancreatic islets.
Heart
Emma
E
BioCurrents Research Center, Marine Biological Laboratory, 7 MBL St., Lillie 219, Woods Hole, MA 02543, USA. eheart@mbl.edu
Cline
Gary W
GW
Collis
Leon P
LP
Pongratz
Rebecca L
RL
Gray
Joshua P
JP
Smith
Peter J S
PJ
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D019343:Citric Acid / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D008293:Malates / Q000378:metabolism*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008928:Mitochondria / Q000378:metabolism*; D010101:Oxygen Consumption / Q000502:physiology; D011766:Pyruvate Carboxylase / Q000378:metabolism*; D051381:Rats; D013386:Succinates / Q000378:metabolism / Q000494:pharmacology
D007328:Insulin; D008293:Malates; D013386:Succinates; D019343:Citric Acid; D000244:Adenosine Diphosphate; C030298:malic acid; D000255:Adenosine Triphosphate; D008291:Malate Dehydrogenase; C030574:malate dehydrogenase (decarboxylating); D011766:Pyruvate Carboxylase; D005947:Glucose; D002118:Calcium; C041105:monomethyl succinate