کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2007478 | 1066376 | 2008 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Characterization of a conformationally sensitive TOAC spin-labeled substance P
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کلمات کلیدی
ESRBCATFAGPCRNK1RTBSGFPFMOCBBA9-FluorenylmethyloxycarbonylMBBFBSTOACBSA - BSAG-protein coupled receptor - G-پروتئین همراه گیرندهGpp(NH)p - GPP (NH) صbovine serum albumin - آلبومین سرم گاوTrifluoroacetic acid - اسید TrifluoroaceticCho - برایbicinchoninic acid - بیسینکنینیک اسیدTris-buffered saline - تریس بافر شورEPR - تشدید پارامغناطیس الکترونfetal bovine serum - سرم جنین گاوChinese hamster ovary cells - سلول های تخمدان هامستر چینیelectron paramagnetic resonance spectroscopy - طیف سنجی رزونانس پارامغناطیسی الکترونSubstance P - ماده Pgreen fluorescent protein - پروتئین فلورسنت سبزhigh performance liquid chromatography - کروماتوگرافی مایع با کارایی بالاHPLC - کروماتوگرافی مایعی کاراNeurokinin-1 receptor - گیرنده نوروکینین-1
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Characterization of a conformationally sensitive TOAC spin-labeled substance P Characterization of a conformationally sensitive TOAC spin-labeled substance P](/preview/png/2007478.png)
چکیده انگلیسی
To probe the binding of a peptide agonist to a G-protein coupled receptor in native membranes, the spin-labeled amino acid analogue 4-amino-4-carboxy-2,2,6,6-tetramethylpiperidino-1-oxyl (TOAC) was substituted at either position 4 or 9 within the substance P peptide (RPKPQQFFGLM-NH2), a potent agonist of the neurokinin-1 receptor. The affinity of the 4-TOAC analog is comparable to the native peptide while the affinity of the 9-TOAC derivative is â¼250-fold lower. Both peptides activate receptor signaling, though the potency of the 9-TOAC peptide is substantially lower. The utility of these modified ligands for reporting conformational dynamics during the neurokinin-1 receptor activation was explored using EPR spectroscopy, which can determine the real-time dynamics of the TOAC nitroxides in solution. While the binding of both the 4-TOAC substance P and 9-TOAC substance P peptides to isolated cell membranes containing the neurokinin-1 receptor is detected, a bound signal for the 9-TOAC peptide is only obtained under conditions that maintain the receptor in its high-affinity binding state. In contrast, 4-TOAC substance P binding is observed by solution EPR under both low- and high-affinity receptor states, with evidence of a more strongly immobilized peptide in the presence of GDP. In addition, to better understand the conformational consequences of TOAC substitution into substance P as it relates to receptor binding and activation, atomistic models for both the 4- and 9-TOAC versions of the peptide were constructed, and the molecular dynamics calculated via simulated annealing to explore the influence of the TOAC substitutions on backbone structure.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Peptides - Volume 29, Issue 11, November 2008, Pages 1919-1929
Journal: Peptides - Volume 29, Issue 11, November 2008, Pages 1919-1929
نویسندگان
Aaron M. Shafer, Clovis R. Nakaie, Xavier Deupi, Vicki J. Bennett, John C. Voss,