کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10537514 | 962757 | 2016 | 44 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Structural and functional consequences of chaperone site deletion in αA-crystallin
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کلمات کلیدی
DLSARPE-19ChaperoneADHDMEMFBSDulbecco's modified Eagle Medium - Eagle Medium اصلاح شده DulbeccoEDTA - اتیلن دی آمین تترا استیک اسید Ethylenediaminetetraacetic acid - اتیلینیدامین تتراستیک اسیدUltraviolet - اشعه فرابنفشAlcohol dehydrogenase - الکل دهیدروژنازTem - این استAggregation - تجمعMolar mass - توده مولارdeletion - حذفApoptosis - خزان یاختهایcircular dichroism - رنگ تابی دورانیStructure-function - ساختار-عملکردfetal bovine serum - سرم جنین گاوCitrate synthase - سیترات سیتواستاتHydrodynamic radius - شعاع هیدرودینامیکیRefractive index - ضریب شکستMALS - مالشTransmission electron microscopy - میکروسکوپ الکترونی عبوریHLE - هلیDynamic Light Scattering - پراکندگی نور دینامیکیmulti-angle light scattering - پراکندگی نور چند زاویهhigh-pressure liquid chromatography - کروماتوگرافی مایع با فشار بالاHPLC - کروماتوگرافی مایعی کاراcrystallin - کریستالین
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The chaperone-like activity of αA-crystallin has an important role in maintaining lens transparency. Previously we identified residues 70-88 as a chaperone site in αA-crystallin. In this study, we deleted the chaperone site residues to generate αAÎ70-76 and αAÎ70-88 mutants and investigated if there are additional substrate-binding sites in αA-crystallin. Both mutant proteins when expressed in E. coli formed inclusion bodies, and on solubilizing and refolding, they exhibited similar structural properties, with a 2- to 3-fold increase in molar mass compared to the molar mass of wild-type protein. The deletion mutants were less stable than the wild-type αA-crystallin. Functionally αAÎ70-88 was completely inactive as a chaperone, while αAÎ70-76 demonstrated a 40-50% reduction in anti-aggregation activity against alcohol dehydrogenase (ADH). Deletion of residues 70-88 abolished the ADH binding sites in αA-crystallin at physiological temperature. At 45 °C, cryptic ADH binding site(s) became exposed, which contributed subtly to the chaperone-like activity of αAÎ70-88. Both of the deletion mutants were completely inactive in suppressing aggregation of βL-crystallin at 53 °C. The mutants completely lost the anti-apoptotic property that αA-crystallin exhibits while they protected ARPE-19 (a human retinal pigment epithelial cell line) and primary human primary lens epithelial (HLE) cells from oxidative stress. Our studies demonstrate that residues 70-88 in αA-crystallin act as a primary substrate binding site and account for the bulk of the total chaperone activity. The β3 and β4 strands in αA-crystallin comprising 70-88 residues play an important role in maintenance of the structure and in preventing aggregation of denaturing proteins.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1864, Issue 11, November 2016, Pages 1529-1538
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1864, Issue 11, November 2016, Pages 1529-1538
نویسندگان
Puttur Santhoshkumar, Srabani Karmakar, Krishna K. Sharma,