Title of article :
Mitochondrial malic enzyme (ME2) in pancreatic islets of the human, rat and mouse and clonal insulinoma cells: Simple enzyme assay for mitochondrial malic enzyme 2
Author/Authors :
MacDonald، نويسنده , , Michael J. and Longacre، نويسنده , , Melissa J. and Kendrick، نويسنده , , Mindy A.، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2009
Pages :
5
From page :
100
To page :
104
Abstract :
Despite interest in malic enzyme(ME)s in insulin cells, mitochondrial malic enzyme (ME2) has only been studied with estimates of mRNA or with mRNA knockdown. Because an mRNA’s level does not necessarily reflect the level of its cognate enzyme, we designed a simple spectrophotometric enzyme assay to measure ME2 activity of insulin cells by utilizing the distinct kinetic properties of ME2. Mitochondrial ME2 uses either NAD or NADP as a cofactor, has a high Km for malate and is allosterically activated by fumarate and inhibited by ATP. Cytosolic ME (ME1) and the other mitochondrial ME (ME3) use only NADP as a cofactor and have lower Kms for malate. The assay easily showed for the first time that substantial ME2 activity is present in pancreatic islets of humans, rats and mice and INS-1 832/13 cells. ME2’s presence was confirmed with immunoblotting. There was no evidence that ME3 is present in these tissues.
Keywords :
Mitochondrial NAD(P) malic enzyme , ME2 , Fumarate activation , malate , ATP inhibition , Pancreatic islets , Spectrophotometric enzyme assay procedure
Journal title :
Archives of Biochemistry and Biophysics
Serial Year :
2009
Journal title :
Archives of Biochemistry and Biophysics
Record number :
1630732
Link To Document :
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