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Recombinant Human Malic Enzyme 2, His

Product Specifications

Background

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.

Product Name Alternative

NAD-dependent Malic Enzyme, Mitochondrial, NAD-ME

Gene ID

4200

UniProt

P23368

Target

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

Purification

> 95 % by SDS-PAGE and HPLC analyses.

Endotoxin

Less than 1 EU/ugof rHuME2, His as determined by LAL method.

Bioactivity

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.

Storage Conditions

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.

Calculated Molecular Weight

Approximately 64.4 kDa, a single non-glycosylated polypeptide chain containing 573 amino acids with 6 x His at C-terminus.

Formulation

Lyophilized from a 0.2 μm filtered concentrated solution in PBS, pH 7.4.

Host or Source

Escherichia coli

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