PYCR1 is a universal housekeeping enzyme that catalyzes the NAD(P)H-dependent conversion of pyrroline-5-carboxylate to proline. This enzyme may also play a physiologic role in the generation of NADP(+) in some cell types. It forms a homopolymer and localizes to the mitochondrion. Defects in PYCR1 are the cause of cutis laxa autosomal recessive type 2B (ARCL2B). Recombinant protein corresponding to aa1-319 of human PYCR1, fused to His-tag at N-terminus, expressed in E. coli. Accession: NP_008838. Molecular Weight: ~35.5kD (339aa), confirmed by MALDI-TOF Amino Acid Sequence: MGSSHHHHHH SSGLVPRGSH MSVGFIGAGQ LAFALAKGFT AAGVLAAHKI MASSPDMDLA TVSALRKMGV KLTPHNKETV QHSDVLFLAV KPHIIPFILD EIGADIEDRH IVVSCAAGVT ISSIEKKLSA FRPAPRVIRC MTNTPVVVRE GATVYATGTH AQVEDGRLME QLLSSVGFCT EVEEDLIDAV TGLSGSGPAY AFTALDALAD GGVKMGLPRR LAVRLGAQAL LGAAKMLLHS EQHPGQLKDN VSSPGGATIH ALHVLESGGF RSLLINAVEA SCIRTRELQS MADQEQVSPA AIKKTILDKV KLDSPAGTAL SPSGHTKLLP RSLAPAGKD Specific Activity: As Reported Biological Activity: One unit will oxidize 1.0umole of L-proline to 1-pyrroline-5-carboxylate per minute in the presence of beta NAD at pH 10.0 at 25C. Activity Assay: 1. Prepare an assay buffer with the following concentrations: 200mM Tris-HCl, pH 10.0 and 10mM NAD, 20mM L-proline, and load 150ul into each well. 2. Add 50ul of 1.25ug and 2.5ug of P9399-25 to each well. 3. Record the increase in A340nm for 5 minutes at 25C. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. 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.
