| Source: |
Escherichia coli |
| Molecular Weight: |
Approximately 64.4 kDa, a single non-glycosylated polypeptide chain containing 573 amino acids with 6 × His at C-terminus. |
| AA Sequence: |
MLHIKEKGKP LMLNPRTNKG MAFTLQERQM LGLQGLLPPK IETQDIQALR FHRNLKKMTS PLEKYIYIMG IQERNEKLFY RILQDDIESL MPIVYTPTVG LACSQYGHIF RRPKGLFISI SDRGHVRSIV DNWPENHVKA VVVTDGERIL GLGDLGVYGM GIPVGKLCLY TACAGIRPDR CLPVCIDVGT DNIALLKDPF YMGLYQKRDR TQQYDDLIDE FMKAITDRYG RNTLIQFEDF GNHNAFRFLR KYREKYCTFN DDIQGTAAVA LAGLLAAQKV ISKPISEHKI LFLGAGEAAL GIANLIVMSM VENGLSEQEA QKKIWMFDKY GLLVKGRKAK IDSYQEPFTH SAPESIPDTF EDAVNILKPS TIIGVAGAGR LFTPDVIRAM ASINERPVIF ALSNPTAQAE CTAEEAYTLT EGRCLFASGS PFGPVKLTDG RVFTPGQGNN VYIFPGVALA VILCNTRHIS DSVFLEAAKA LTSQLTDEEL AQGRLYPPLA NIQEVSINIA IKVTEYLYAN KMAFRYPEPE DKAKYVKERT WRSEYDSLLP DVYEWPESAS SPPVITEHHH HHH |
| Purity: |
> 95% by SDS-PAGE and HPLC analyses. |
| Biological 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. |
| Physical Appearance: |
Sterile filtered white lyophilized (freeze-dried) powder. |
| Formulation: |
Lyophilized from a 0.2 μm filtered concentrated solution in PBS, pH7.4. |
| Endotoxin: |
Less than 1 EU/μg of rHuME2, His as determined by LAL method. |
| 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°C. Further dilutions should be made in appropriate buffered solutions. |
| Stability & Storage: |
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70°C as supplied.
1 month, 2 to 8°C under sterile conditions after reconstitution.
3 months, -20 to -70°C under sterile conditions after reconstitution. |
| Synonyms: |
NAD-dependent Malic Enzyme, Mitochondrial, NAD-ME |
| Background: |
The ME2 gene in humans encodes a mitochondrial NAD-dependent malic enzyme, a homotetrameric protein that catalyzes the oxidative decarboxylation of malate to pyruvate. This enzyme exists in three isoforms within mammalian tissues: a strictly cytosolic NADP+-dependent form, an NADP+-dependent mitochondrial isoform, and a mitochondrial isoenzyme capable of utilizing both NAD+ and NADP+, with a preference for NAD+. The mammalian isoforms typically weigh approximately 62-64 kDa, suggesting a tetrameric structure for the active enzyme, as indicated by its native size of 240,000 Da. |
