کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2599768 1562636 2012 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Polyethylene glycosylation prolongs the stability of recombinant human paraoxonase-1
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم محیط زیست بهداشت، سم شناسی و جهش زایی
پیش نمایش صفحه اول مقاله
Polyethylene glycosylation prolongs the stability of recombinant human paraoxonase-1
چکیده انگلیسی

Paraoxonase-1 (PON1) is a native enzyme that is synthesized in the liver and is capable of hydrolyzing organophosphates (OPs). It is regarded as part of a promising approach for the pretreatment and therapy of OP poisoning. Previous experiments with purified rabbit serum PON1 have established that it can protect rats against many OP exposures. In the current paper, we described a preparation of active recombinant human PON1 (rHuPON1) by engineering an Escherichia coli expression system. Recombinant HuPON1 was purified by Ni-NTA affinity chromatography followed by DEAE sepharose fast-flow chromatography. After purification, rHuPON1 was chemically modified with polyethyleneglycol (PEG)-20K. Recombinant HuPON1 exhibited a mean residence time (MRT) of 8.9 h, which was threefold shorter than that of native HuPON1 in rats. However, rHuPON1 chemically modified with PEG-20K displayed an MRT of 19.5 h, suggesting that PEG modification can prolong the circulatory stability of rHuPON1. PEG-rHuPON1 had a catalytic efficiency sufficient in protecting rats against OP poisoning, as measured by acetylcholinesterase activity in tissues and signs after poisoning.


► Active recombinant human PON1 (rHuPON1) was engineered in an Escherichia coli expression system.
► Polyethylene glycol-20K modification was used to prolong rHuPON1 residence time.
► Polyethylene glycosylation prolonged the mean residence time of rHuPON1 from 8.9 h to19.5 h.
► Modified rHuPON1 had sufficient protection against organophosphate exposure in rats.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Toxicology Letters - Volume 210, Issue 3, 5 May 2012, Pages 366–371
نویسندگان
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