کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10537687 | 962814 | 2005 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Preparation and characterization of a single-chain calcineurin-calmodulin complex
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کلمات کلیدی
CNAFPLCCBACNBpNPPp-nitrophenyl phosphate - p-نیترفنیل فسفاتInteraction - اثر متقابلcircular dichroism - رنگ تابی دورانیCAM - ساخت به کمک کامپیوترfast protein liquid chromatography - سریع کروماتوگرافی مایع پروتئینPhosphatase activity - فعالیت فسفاتازFusion - فیوژنCalmodulin - کالمودولینCalcineurin - کلسینورین
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Preparation and characterization of a single-chain calcineurin-calmodulin complex Preparation and characterization of a single-chain calcineurin-calmodulin complex](/preview/png/10537687.png)
چکیده انگلیسی
Calcineurin (CN), a Ca2+/calmodulin (CaM)-dependent serine/threonine protein phosphatase, is a heterodimer composed of a catalytic subunit (CNA) and a regulatory subunit (CNB). The activity of CNA is under the control of two functionally distinct, but structurally similar Ca2+-regulated proteins, CaM and CNB. The crystal structure of the holoenzyme reveals that the N-terminus and C-terminus of CNB and the N-terminus of CNA each have a long arm not involved in the active site. We constructed a fusion of the genes of CaM, CNB and CNA in that order using linker primers containing six and ten codons of glycine. A single-chain CaM-CNB-CNA (CBA) complex was expressed and purified to near homogeneity. The single-chain complex was fully soluble, and had biochemical properties and kinetic parameters similar to single-chain CNB-CNA (BA) activated by CaM. It was not regulated by CaM and CNB, but was strongly stimulated by Mn2+, Ni2+ and Mg2+. Intrinsic fluorescence spectroscopy of the complex showed a change in the environment of tryptophan in the presence of Ca2+ and circular dichroism (CD) spectropolarimetry revealed an increase in alpha-helical content. Our findings suggest that fusion of CaM, CNB and CNA does not prevent the structural changes required for their functioning; in particular, CaM within the complex could still interact correctly with CN in the presence of Ca2+.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1747, Issue 2, 14 March 2005, Pages 171-178
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1747, Issue 2, 14 March 2005, Pages 171-178
نویسندگان
Yunlong Qin, Jing Liu, Xin Li, Qun Wei,