کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
10532582 961630 2013 29 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Correction factors for membrane protein molecular weight readouts on sodium dodecyl sulfate-polyacrylamide gel electrophoresis
ترجمه فارسی عنوان
فاکتورهای اصلاح شده برای اندازه گیری وزن مولکول پروتئین غشایی بر روی الکتروفورز ژل دوده سولفات سدیم پلی آکریل آمید
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی
Membrane proteins are known to migrate anomalously on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to relative molecular mass (Mr) values larger or smaller than formula molecular weight. We constructed a database from literature Mr values reported for 168 nonredundant helical membrane proteins with structures determined to high resolution and found that more than three-quarters of them exhibit this behavior on gels calibrated with commercial standards. Further analysis of the database indicated that the direction of anomalous migration is not a consequence of membrane protein net charge or hydrophobicity. Plots of observed versus formula Mr values showed that membrane proteins migrating slower than expected read out at 1.13 × Mr, whereas those that migrate faster than expected read out at 0.82 × Mr (R2 ∼ 0.98, P < 0.0001). These robust trends imply that division of the Mr readouts of slower or faster migrating analytes by 1.13 or 0.82, respectively, should enhance SDS-PAGE accuracy. Applying this correction procedure to SDS-PAGE readouts of four fast-migrating helical transmembrane (TM) proteins significantly reduced Mr errors from approximately 20% to 8% (P < 0.0001). Our results suggest that hydrophobic standards for SDS-PAGE would significantly improve the performance of the technique applied to membrane proteins.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 434, Issue 1, 1 March 2013, Pages 67-72
نویسندگان
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