||Recombinant human PPIE was expressed in E.coli and purified by conventional chromatography techniques, 37.5 kDa (337 aa).
||Cyclophilins possess the activity of peptidyl-prolyl cis-trans isomerase (PPIase) and are involved in cellular protein folding and protein interactions. PPIE contains two RNA binding domains at the N-terminal region and a PPIase domain at the C-terminal region. It was not known what cellular and physiological roles of PPIE.
|Sequences of amino acids:
||MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDRWGSMATT KRVLYVGGLA EEVDDKVLHA AFIPFGDITD IQIPLDYETE KHRGFAFVEF ELAEDAAAAI DNMNESELFG RTIRVNLAKP MRIKEGSSRP VWSDDDWLKK FSGKTLEENK EEEGSEPPKA ETQEGEPIAK KARSNPQVYM DIKIGNKPAG RIQMLLRSDV VPMTAENFRC LCTHEKGFGF KGSSFHRIIP QFMCQGGDFT NHNGTGGKSI YGKKFDDENF ILKHTGPGLL SMANSGPNTN GSQFFLTCDK TDWLDGKHVV FGEVTEGLDV LRQIEAQGSK DGKPKQKVII ADCGEYV
||Liquid. In 20 mM Tris-HCl buffer (pH 8.0)
||> 95% by SDS - PAGE
||1 mg/ml (determined by Bradford assay)
||Specific activity is > 210 nmoles/min/ug, and is defined as the amount of enzyme that cleave 1umole of suc-AAFP-pNA per minute at 1°C in Tris-HCl pH8.0 using chymotrypsin.
||1. Prepare assay buffer into a suitable container and pre-chilling on ice before use: the final concentrations are 35 mM Tris, pH 8.0, 1uM suc-AAFP-pNA). 2. Add recombinant Cyclophilin E protein with various concentrations (1ug, 2ug, 5ug) in assay buffer. 3. Mix by inversion and equilibrate to 1°C and monitor the A410nm until the value is constant using a spectrophotometer. 4. Add pre-chilled chymotrypsin to 12.5 uM and mix immediately. 5. Record the increase in A410 nm for 20 minutes.
||Can be stored at +4°C short term (1-2 weeks). For long term storage, aliquot and store at -20°C or -70°C. Avoid repeated freezing and thawing cycles.