کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8941290 | 1644994 | 2018 | 35 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Generating a recombinant phosphothreonine-binding domain for a phosphopeptide of the human transcription factor, c-Myc
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کلمات کلیدی
Fbxw7CDKFN3mAbMYCPTBDSCFERKPP2AGSK3βPABCaMKIIMonoclonal antibody - آنتی بادی مونوکلونالPolyclonal antibody - آنتی بادی های پلی کلونالFibronectin type III domain - دامنه فیبرونکتین نوع IIIphosphothreonine - فسفاترونینPhosphorylation - فسفریلاسیونphosphoserine - فسفسئینPhage display - نمایش فاژیcalmodulin-dependent protein kinase II - وابسته به کالدولین پروتئین کیناز IIAntibody - پادتَن یا آنتیبادیprotein phosphatase 2A - پروتئین فسفاتاز 2Aextracellular signal-regulated kinase - کیناز تنظیم شده سیگنال خارج سلولیcyclin-dependent kinase - کییناز وابسته به سیکلینGlycogen synthase kinase-3β - گلیکوزین سنتاز کیناز 3β
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
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چکیده انگلیسی
Transcription factor c-Myc is an oncoprotein that is regulated at the post-translational level through phosphorylation of two conserved residues, Serine 62 (Ser62) and Threonine 58 (Thr58). A highly specific tool capable of recognizing Myc via pThr58 is needed to monitor activation and localization. Through phage display, we have isolated 10 engineered Forkhead-associated (FHA) domains that selectively bind to a phosphothreonine (pThr)-containing peptide (53-FELLPpTPPLSPS-64) segment of human c-Myc. One domain variant was observed to bind to the Myc-pThr58 peptide with a KD value of 800â¯nM and had >1000-fold discrimination between the phosphorylated and non-phosphorylated peptide. The crystal structure of the engineered FHA Myc-pThr-binding domain (Myc-pTBD) was solved in complex with its cognate ligand. The Myc-pTBD was observed to be structurally similar to the yeast Rad9 FHA1 domain, except that its β4-β5 and β10-β11 loops form a hydrophobic pocket to facilitate the interaction between the domain and the peptide ligand. The Myc-pTBD's specificity for its cognate ligand was demonstrated to be on a par with 3 commercial polyclonal antibodies, suggesting that this recombinant reagent is a viable alternative to antibodies for monitoring Myc regulation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: New Biotechnology - Volume 45, 25 October 2018, Pages 36-44
Journal: New Biotechnology - Volume 45, 25 October 2018, Pages 36-44
نویسندگان
Leon A. Venegas, Stefanie L. Kall, Oluwadamilola Bankole, Arnon Lavie, Brian K. Kay,