[Expression and characterization of protein disulfide isomerase-related protein A].
Protein disulfide isomerase-related protein A (PRPA) was highly expressed (about 34%) in Escherichia coli by inserting the whole PRPA cDNA into the vector pET23b. After expression, the purified protein was acquired through ammonium fractional precipitation and Bio-Rex 70 chromatography. PRPA shows low disulfide isomerase activity (only about 1/250 of that of hPDI), decreases the reactivation yield of denatured and reduced lysozyme either in redox and non-redox Hepes buffer or redox PBS buffer and facilitates the aggregation of denatured and reduced lysozyme. Fluorescence spectra of PRPA indicate that PRPA has more hydrophobic groups at surface than that of hPDI, and which can be used to explain why PRPA has anti-chaperone activity during the refolding of denatured and reduced lysozyme.
Zhou
Hai-Ping
HP
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Jia
Cui-Juan
CJ
Zhang
Yu-Ying
YY
D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D009113:Muramidase / Q000737:chemistry; D010957:Plasmids; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D013050:Spectrometry, Fluorescence
D005656:Fungal Proteins; C404324:PRPA protein, Aspergillus niger; D011994:Recombinant Proteins; D009113:Muramidase