کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9885109 | 1537150 | 2005 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Oligomerization of the Saccharomyces cerevisiae Na+/H+ antiporter Nha1p: Implications for its antiporter activity
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
The Na+/H+ antiporter (Nha1p) from the budding yeast Saccharomyces cerevisiae plays an important role in intracellular pH and Na+ homeostasis. Here, we show by co-precipitation of differently tagged Nha1p proteins expressed in the same cell that the yeast Nha1p l forms an oligomer. In vitro cross-linking experiments then revealed that Nha1p-FLAG is present in the membranes as a dimer. Differently tagged Nha1p proteins were also co-precipitated from sec18-1 mutant cells in which ER-to-Golgi traffic is blocked under non-permissive temperatures, suggesting that Nha1p may already dimerize in the ER membrane. When we over-expressed a mutant Nha1p with defective antiporter activity in cells that also express the wild-type Nha1p-EGFP fusion protein, we found impaired cell growth in highly saline conditions, even though the wild-type protein was appropriately expressed and localized correctly. Co-immunoprecipitation assays then showed the inactive Nha1p-FLAG mutant interacted with the wild-type Nha1p-EGFP protein. These results support the notion that Nha1p exists in membranes as a dimer and that the interaction of its monomers is important for its antiporter activity.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Biomembranes - Volume 1720, Issues 1â2, 30 December 2005, Pages 125-136
Journal: Biochimica et Biophysica Acta (BBA) - Biomembranes - Volume 1720, Issues 1â2, 30 December 2005, Pages 125-136
نویسندگان
Keiji Mitsui, Hidetomo Yasui, Norihiro Nakamura, Hiroshi Kanazawa,