کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8359366 | 1542291 | 2018 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Solubilization conditions for bovine heart mitochondrial membranes allow selective purification of large quantities of respiratory complexes I, III, and V
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کلمات کلیدی
NADH dehydrogenaseQ10DTTFMNEDTA - اتیلن دی آمین تترا استیک اسید Ethylenediaminetetraacetic acid - اتیلینیدامین تتراستیک اسیدblue native polyacrylamide gel electrophoresis - الکتروفورز ژل پلی آکریل آمید آبی بومیMitochondrial respiration - تنفس میتوکندریdithiothreitol - دیتیوتریتولsuccinate dehydrogenase - سوکسیناد دهیدروژنازcytochrome c oxidase - سیتوکروم سی اکسیدازphosphatidylcholine - فسفاتیدیل کولینphosphatidylethanolamine - فسفاتیدیلتانولامینflavin mononucleotide - فلاون مونونوکلئوتیدComplex I - مجتمع IComplex IV - مجتمع IVComplex V - مجتمع VComplex II - مجتمع دومPurification - پاکسازیcomplex III - پیچیده IIICardiolipin - کاردیولیپین
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Ascertaining the structure and functions of mitochondrial respiratory chain complexes is essential to understanding the biological mechanisms of energy conversion; therefore, numerous studies have examined these complexes. A fundamental part of that research involves devising a method for purifying samples with good reproducibility; the samples obtained need to be stable and their constituents need to retain the same structure and functions they possess when in mitochondrial membranes. Submitochondrial bovine heart particles were isolated using differential centrifugation to adjust to a membrane concentration of 46.0% (w/v) or 31.5% (w/v) based on weight. After 0.7% (w/v) deoxycholic acid, 0.4% (w/v) decyl maltoside, and 7.2% (w/v) potassium chloride were added to the mitochondrial membranes, those membranes were solubilized. At a membrane concentration of 46%, complex V was selectively solubilized, whereas at a concentration of 31.5% (w/v), complexes I and III were solubilized. Two steps-sucrose density gradient centrifugation and anion-exchange chromatography on a POROS HQ 20â¯Î¼m column-enabled selective purification of samples that retained their structure and functions. These two steps enabled complexes I, III, and V to be purified in two days with a high yield. Complexes I, III, and V were stabilized with n-decyl-β-D-maltoside. A total of 200â¯mg-300â¯mg of those complexes from one bovine heart (1.1â¯kg muscle) was purified with good reproducibility, and the complexes retained the same functions they possessed while in mitochondrial membranes.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 150, October 2018, Pages 33-43
Journal: Protein Expression and Purification - Volume 150, October 2018, Pages 33-43
نویسندگان
Satoru Shimada, Shintaro Maeda, Masahide Hikita, Kaoru Mieda-Higa, Shigefumi Uene, Yukiko Nariai, Kyoko Shinzawa-Itoh,