کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10747962 | 1050254 | 2016 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Ca2+/calmodulin-dependent protein kinase phosphatase (CaMKP/PPM1F) interacts with neurofilament L and inhibits its filament association
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کلمات کلیدی
CaMKPDMEMDTT2-MENFLGAPDHNGFCaMKPMSFCBB2-mercaptoethanol - 2-мерکارپتواتانولBSA - BSACoomassie Brilliant Blue - Coomassie درخشان آبیbovine serum albumin - آلبومین سرم گاوInteraction analysis - تجزیه و تحلیل تعاملdithiothreitol - دیتیوتریتولintermediate filament - رشته های متوسطMass spectrometry - طیف سنجی جرمیnerve growth factor - فاکتور رشد عصبphenylmethylsulfonyl fluoride - فنیل متیل سولفونیل فلورایدDulbecco’s modified eagle’s medium - محیط عقاب اصلاح شده DulbeccoModulator - مدولاتورNeuron - نورونCa2+/calmodulin-dependent protein kinase - پروتئین کیناز وابسته به کلسیم و پتاسیم Ca2 + /glyceraldehyde-3-phosphate dehydrogenase - گلیسرالیدید-3-فسفات دهیدروژناز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
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چکیده انگلیسی
Ca2+/calmodulin-dependent protein kinase phosphatase (CaMKP/PPM1F) is a Ser/Thr phosphatase that belongs to the PPM family. Growing evidence suggests that PPM phosphatases including CaMKP act as a complex with other proteins to regulate cellular functions. In this study, using the two-dimensional far-western blotting technique with digoxigenin-labeled CaMKP as a probe, in conjunction with peptide mass fingerprinting analysis, we identified neurofilament L (NFL) as a CaMKP-binding protein in a Triton-insoluble fraction of rat brain. We confirmed binding of fluorescein-labeled CaMKP (F-CaMKP) to NFL in solution by fluorescence polarization. The analysis showed that the dissociation constant of F-CaMKP for NFL is 73 ± 17 nM (n = 3). Co-immunoprecipitation assay using a cytosolic fraction of NGF-differentiated PC12 cells showed that endogenous CaMKP and NFL form a complex in cells. Furthermore, the effect of CaMKP on self-assembly of NFL was examined. Electron microscopy revealed that CaMKP markedly prevented NFL from forming large filamentous aggregates, suggesting that CaMKP-binding to NFL inhibits its filament association. These findings may provide new insights into a novel mechanism for regulating network formation of neurofilaments during neuronal differentiation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 477, Issue 4, 2 September 2016, Pages 820-825
Journal: Biochemical and Biophysical Research Communications - Volume 477, Issue 4, 2 September 2016, Pages 820-825
نویسندگان
Hana Ozaki, Tsuyoshi Katoh, Ryoko Nakagawa, Yasuhiro Ishihara, Noriyuki Sueyoshi, Isamu Kameshita, Takanobu Taniguchi, Tetsuo Hirano, Takeshi Yamazaki, Atsuhiko Ishida,