کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10825176 | 1063355 | 2005 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Reconstitution of glycopeptide export in mixed detergent-solubilised and resealed microsomes depleted of lumenal components
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کلمات کلیدی
PDIConcanavalin APMSFConARibophorin Isignal recognition particle receptorReconstitution - بازسازیRetrograde transport - حمل بار عقبگردendoplasmic reticulum - شبکه آندوپلاسمی phenylmethylsulphonyl fluoride - فنیل متیل سولفونیل فلورایدProtein disulphide isomerase - پروتئین دی سولفید ایزومرازquality control - کنترل کیفیتGlycopeptide - گلیکوپپتید
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
Export of macromolecules from the endoplasmic reticulum (ER) lumen into the cytosol is a major aspect of the quality control systems operating within the early secretory system. Glycopeptides are exported from the ER by an ATP- and GTP-dependent pathway, which shares many similarities to the protein export system. Significantly, for glycopeptides, there is no requirement for cytosolic factors, biochemically distinguishing the glycopeptide and protein paths and probably reflecting the lower conformational complexity of the former substrate. Genetic studies in yeast, and biochemical data from higher eukaryotes, indicate that glycopeptides utilise the Sec61 translocon. Here, we report a new system allowing access to lumenal ER components, facilitating assessment of their importance in glycopeptide retrotranslocation and potentially other processes. Saponin, in combination with CHAPS, but not saponin alone, facilitated removal of >95% of lumenal protein disulphide isomerase (PDI) and BiP. Upon resealing, these microsomes retained glycopeptide export competence. These data suggest that the majority of lumenal components of the ER are most likely nonessential for glycopeptide export. In addition, export competence was highly sensitive to the addition of external protease, indicating a role for protein factors with cytoplasmically exposed determinants.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biochemical and Biophysical Methods - Volume 62, Issue 1, 31 January 2005, Pages 1-12
Journal: Journal of Biochemical and Biophysical Methods - Volume 62, Issue 1, 31 January 2005, Pages 1-12
نویسندگان
Bassam R.S. Ali, Laura C. Edwards, Mark C. Field,