Abstract:
:Human phospholipid scramblase 1 (hPLSCR1) scrambles plasma membrane phospholipids during cell activation, blood coagulation and apoptosis. It was over-expressed in E. coli with a histidine tag and purified from the inclusion bodies (*30 mg/l culture broth) under denaturing conditions using 8 M urea. The denatured hPLSCR1 refolded into its native configuration when urea was removed as shown by a 10-fold increase in its intrinsic fluorescence. Active hPLSCR1 showed scrambling activity in vitro after reconstituting in proteoliposomes. hPLSCR1 showed higher rates of scrambling activity for phosphatidylethanolamine than phosphatidylcholine. Binding studies with the calcium analogue "Stains-all" dye showed a characteristic peak, termed as the J band, at 650 nm. This is the first report on high level expression of hPLSCR1 with histidine tag in E. coli.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Sahu SK,Gopala Krishna A,Gummadi SNdoi
10.1007/s10529-008-9797-zsubject
Has Abstractpub_date
2008-12-01 00:00:00pages
2131-7issue
12eissn
0141-5492issn
1573-6776journal_volume
30pub_type
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