Description : |
NAD-dependent malic enzyme (ME2), mitochondrial is a protein that in humans is encoded by the ME2 gene. This gene encodes a mitochondrial NAD-dependent malic enzyme, a homotetrameric protein, which catalyzes the oxidative decarboxylation of malate to pyruvate. Three different isoforms of ME are known to be in mammalian tissues: a strictly cytosolic NADP+-dependent enzyme, an NADP+-dependent mitochondriail isoform, and a mitochondrial isoenzyme that is able to use both NAD+ and NADP+ but is more effective with NAD+. The mammalian isoforms size is about 62-64 kDa. A native size of 240,000 Da proposes a tetrameric structure for the active enzyme. |
Source : |
E.coli |
Species : |
Human |
Form : |
Lyophilized from a 0.2μm filtered concentrated solution in PBS, pH 7.4. |
Bio-activity : |
Malic Enzyme activity was assayed spectrophotometrically at 340nm as described in Mandela and Sauer (1975). The standard reaction mixture contained 50 mM Tris-HCl, 3 mM MnCl2, 5 mM malate, 0.12 mM NADP+, 2.5 mM fumarate. Assay was performed in a Beckman spectrophotometer. The Km value is 1.5 ± 0.6 mM. |
Molecular Mass : |
Approximately 64.4 kDa, a single non-glycosylated polypeptide chain containing 573 amino acids with 6 × His at C-terminus. |
Protein length : |
573 |
AA Sequence : |
MLHIKEKGKPLMLNPRTNKGMAFTL QERQMLGLQGLLPPKIETQDIQALR FHRNLKKMTSPLEKYIYIMGIQERN EKLFYRILQDDIESLMPIVYTPTVG LACSQYGHIFRRPKGLFISISDRGH VRSIVDNWPENHVKAVVVTDGERIL GLGDLGVYGMGIPVGKLCLYTACAG IRPDRCLPVCIDVGTDNIALLKDPF YMGLYQKRDRTQQYDDLIDEFMKAI TDRYGRNTLIQFEDFGNHNAFRFLR KYREKYCTFNDDIQGTAAVALAGLL AAQKVISKPISEHKILFLGAGEAAL GIANLIVMSMVENGLSEQEAQKKIW MFDKYGLLVKGRKAKIDSYQEPFTH SAPESIPDTFEDAVNILKPSTIIGV AGAGRLFTPDVIRAMASINERPVIF ALSNPTAQAECTAEEAYTLTEGRCL FASGSPFGPVKLTDGRVFTPGQGNN VYIFPGVALAVILCNTRHISDSVFL EAAKALTSQLTDEELAQGRLYPPLA NIQEVSINIAIKVTEYLYANKMAFR YPEPEDKAKYVKERTWRSEYDSLLP DVYEWPESASSPPVITEHHHHHH |
Endotoxin : |
Less than 1 EU/μg of rHuME2, His as determined by LAL method. |
Purity : |
>95% by SDS-PAGE and HPLC analysis. |
Storage : |
Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 centigrade as supplied. 1 month, 2 to 8 centigrade under sterile conditions after reconstitution. 3 months, -20 to -70 centigrade under sterile conditions after reconstitution. |
Reconstitution : |
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1 % BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤-20 centigrade. Further dilutions should be made in appropriate buffered solutions. |