کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10751646 | 1050317 | 2015 | 28 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Cavitation during the protein misfolding cyclic amplification (PMCA) method - The trigger for de novo prion generation?
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کلمات کلیدی
8OHdGrPrPPrPsc5-(Diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxideDEPMPOPrPresDMPOPrPPMCA5,5-dimethyl-1-pyrroline-N-oxide - 5،5-دی متیل-1-پیرولین-N-اکسیدDe novo - باز همElectron paramagnetic resonance - تشدید پارامغناطیس الکترونEPR - تشدید پارامغناطیس الکترونProtein misfolding cyclic amplification - تقویت سیکل ناپایدار پروتئینCavitation - حفره زایی یا کاویتاسیونFree radical - رادیکالهای آزاد Sonication - سونیکیشن 8-hydroxyguanosine - هیدروکسی گوانوزین 8Prion protein - پروتئین پریونPrion - پریون
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The protein misfolding cyclic amplification (PMCA) technique has become a widely-adopted method for amplifying minute amounts of the infectious conformer of the prion protein (PrP). PMCA involves repeated cycles of 20 kHz sonication and incubation, during which the infectious conformer seeds the conversion of normally folded protein by a templating interaction. Recently, it has proved possible to create an infectious PrP conformer without the need for an infectious seed, by including RNA and the phospholipid POPG as essential cofactors during PMCA. The mechanism underpinning this de novo prion formation remains unknown. In this study, we first establish by spin trapping methods that cavitation bubbles formed during PMCA provide a radical-rich environment. Using a substrate preparation comparable to that employed in studies of de novo prion formation, we demonstrate by immuno-spin trapping that PrP- and RNA-centered radicals are generated during sonication, in addition to PrP-RNA cross-links. We further show that serial PMCA produces protease-resistant PrP that is oxidatively modified. We suggest a unique confluence of structural (membrane-mimetic hydrophobic/hydrophilic bubble interface) and chemical (ROS) effects underlie the phenomenon of de novo prion formation by PMCA, and that these effects have meaningful biological counterparts of possible relevance to spontaneous prion formation in vivo.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 461, Issue 3, 5 June 2015, Pages 494-500
Journal: Biochemical and Biophysical Research Communications - Volume 461, Issue 3, 5 June 2015, Pages 494-500
نویسندگان
Cathryn L. Haigh, Simon C. Drew,