کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10841595 | 1067896 | 2005 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Purification and subunit determination of H+-pyrophosphatase from endoplasmic reticulum-enriched vesicles of mung bean seedlings
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کلمات کلیدی
DOCBHTDTTEGTADCCDBSA - BSAN,N′-dicyclohexylcarbodiimide - N، N'-dicyclohexylcarbodiimidebovine serum albumin - آلبومین سرم گاوEDTA - اتیلن دی آمین تترا استیک اسید Deoxycholic acid - اسید Deoxycholicethylenediamine tetraacetic acid - اسید اتیلنیدین تتراستیکbutylhydroxytoluene - بوتیل هیدروکسیتولینdithiothreitol - دیتیوتریتول
موضوعات مرتبط
علوم زیستی و بیوفناوری
علوم کشاورزی و بیولوژیک
دانش گیاه شناسی
پیش نمایش صفحه اول مقاله

چکیده انگلیسی
Endoplasmic reticulum (ER)-enriched vesicles from etiolated hypocotyls of mung bean seedlings (Vigna radiata L.) were isolated by Ficoll gradient and two-polymer phase partition. These ER-enriched vesicles contain a new type of H+-pyrophosphatase (H+-PPase) distinct from that of tonoplasts in higher plants. H+-PPase was then solubilized differentially by deoxycholic acid and lyso-phosphatidylcholine. The solubilized fraction was then subjected to Sephacryl S-200 gel filtration and Mono-Q anion exchange chromatography. The final purified protein complex of ER H+-PPase (ER-PPase) was successfully obtained to high homogeneity. An approximate molecular mass of 170 kDa was determined for the purified ER-PPase by size-exclusion gel filtration chromatography. However, only a single polypeptide of 74 kDa was observed on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Moreover, radiation inactivation analysis of ER-enriched vesicles and purified ER-PPase yielded functional masses of 178.6 ± 9.2 and 143.4 ± 4.7 kDa for inorganic pyrophosphate hydrolysis activity, respectively, indicating that ER-PPase was functionally homodimeric.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Plant Science - Volume 169, Issue 5, November 2005, Pages 847-853
Journal: Plant Science - Volume 169, Issue 5, November 2005, Pages 847-853
نویسندگان
Soong Yu Kuo, Lee Feng Chien, Ru Chun Van, Kun Huang Yan, Pei Feng Liu, Wen Chi Chang, Jung Kai Wang, Rong Long Pan,