Recombinant Human Malic Enzyme 2, His

Synonyms NAD-dependent Malic Enzyme, Mitochondrial, NAD-ME

Accession P23368

GeneID 4200

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.

Quantity 5µg/25µg/1mg

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, pH 7.4.

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.

Usage This material is offered by Shanghai PrimeGene Bio-Tech for research, laboratory or further evaluation purposes. NOT FOR HUMAN USE.

Reference 1. Cousins AB, Badger MR, von Caemmerer S. 2008. J Exp Bot, 59: 1695-703.

2. Murmu JandRaghavendra AS. 2005. J Plant Physiol, 162: 1095-102.

3. Murmu J, Chinthapalli B, Raghavendra AS. 2003. J Exp Bot, 54: 2661-8.

4. Hill S, Winning B, Jenner H, et al. 1996. Biochem Soc Trans, 24: 743-6.

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.