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Updated: Jun 21, 2026

A Fluorescence-based Assay of Phospholipid Scramblase Activity
Published on: September 20, 2016
GroES and GroEL are essential chaperones for refolding of recombinant human phospholipid scramblase 1 in E. coli
Santosh Kumar Sahu1, Archita Rajasekharan, Sathyanarayana N Gummadi
1Applied and Industrial Microbiology Laboratory, Department of Biotechnology, Indian Institute of Technology-Madras, Chennai, 600 036, India.
Abstract:
Human phospholipid scramblase 1(hPLSCR1), when expressed in E. coli (BL-21 DE3), forms inclusion bodies that are functionally inactive. We studied the effects of various stress inducing agents and chaperones on soluble expression of hPLSCR1 in E. coli (BL-21 DE3). Addition of 3% (v/v) ethanol before induction and decreasing the post-induction temperature to 15 degrees C increased the solubility of hPLSCR1 to approximately 10 and approximately 15% respectively. Presence of groES-groEL complex solubilized the hPLSCR1 to approximately 30% of the total hPLSCR1. Absence of groES-groEL did not improve the solubility of hPLSCR1 suggesting that groES and groEL are the essential chaperones for the correct folding of hPLSCR1 when over-expressed in E. coli.
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