NMNAT1, Recombinant, Human, aa115-270, His-Tag (Nicotinamide nucleotide adenylyltransferase 1, NMNAT, PNAT1)

Blue Ice

36

Supplied as a liquid in 20mM Tris-HCl, pH 8.0, 0.1M sodium chloride, 1mM DTT, 1mM EDTA, 20% glycerol.

NMNAT1, also known as NMNAT or PNAT1, is a central enzyme in NAD biosynthesis, catalyzing the condensation of nicotinamide mononucleotide (NMN) or nicotinic acid mononucleotide (NaMN) with the AMP moiety of ATP to form NAD or NaAD. It is widely expressed with high levels in skeletal muscle, heart, liver and kidney. This protein appears to have the ability to protect against axonal degeneration following mechanical or toxic insults.

Source:
Recombinant protein corresponding to aa115-270 from human NMNAT1, fused to His-tag at N-terminal expressed in E. coli.

Molecular Weight:
~36kD(315aa)

Biological Activity:
Specific activity is >7000 pmol/min/ug, and was obtained by measuring the beta-NAD from nicotinamide mononucleotide and ATP/minute at pH 8.0 at 37°C.

Activity Assay:
1. Prepare a 180ul assay buffer into a suitable container. The concentrations are 50mM Tris-HCl (pH 8.0), 50mM semicarbazide, 22mM magnesium chloride (MgCl2), 2.2mM beta-NMN, 2.2mM ATP, 5.5unit ADH, 2.2% (v/v) ethanol.
2. Equilibrate to 37°C and monitor at A340nm until the value is constant using a spectrophotometer.
3. Add 20ul of recombinant NMNAT1 protein 20ug/ml in assay buffer.
4. Mix by inversion and record the decrease at A340nm for approximately 5 minutes.

Amino Acid Sequence:
MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDRWGSMENS EKTEVVLLAC GSFNPITNMH LRLFELAKDY MNGTGRYTVV KGIISPVGDA YKKKGLIPAY HRVIMAELAT KNSKWVEVDT WESLQKEWKE TLKVLRHHQE KLEASDCDHQ QNSPTLERPG RKRKWTETQD SSQKKSLEPK TKAVPKVKLL CGADLLESFA VPNLWKSEDI TQIVANYGLI CVTRAGNDAQ KFIYESDVLW KHRSNIHVVN EWIANDISST KIRRALRRGQ SIRYLVPDLV QEYIEKHNLY SSESEDRNAG VILAPLQRNT AEAK

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.