A New Functional Model for Prediction of Chaperone Activity of the Recombinant M. tb Acr (α-Crystallin) Using Insulin as Substrate
المؤلفون المشاركون
المصدر
Canadian Journal of Infectious Diseases and Medical Microbiology
العدد
المجلد 2019، العدد 2019 (31 ديسمبر/كانون الأول 2019)، ص ص. 1-18، 18ص.
الناشر
Hindawi Publishing Corporation
تاريخ النشر
2019-02-10
دولة النشر
مصر
عدد الصفحات
18
التخصصات الرئيسية
الملخص EN
Mycobacterium tuberculosis Acr is an important protein expressed in latent tuberculosis which is active as an oligomer in preventing misfolding of cellular proteins.
In this study, Mycobacterium alpha crystallin (acr) gene was cloned and expressed in Escherichia coli (E.
coli).
The recombinant Acr protein was purified by Nickel-NTA resin.
The oligomeric state of Acr was confirmed by gel filtration chromatography using Sephacryl S-200 and Native-PAGE.
Studies of chaperone activity were performed with insulin as a substrate at different mole ratios of Acr with 2 types of samples, His tag elutes (H) and His tag elutes with gel filtration (G).
It was observed that the ratio of different sizes of oligomers (9 to 24 mers) had a significant effect on chaperone activity.
Using the mole ratio of Acr for both (H) and (G) samples to insulin B chain and ratio of oligomers, we determined the number of Acr molecules binding to insulin as a model substrate.
We found that if 1.5% of the insulin B chains are covered completely by the (G) samples, aggregation is completely inhibited as compared to 6% with (H) samples.
Pre-heat treatment studies were carried out at 37°C, 60°C, and 70°C.
Far-ultraviolet Circular Dichroism (UV-CD) analysis provided fresh insights into the role of β-sheets and α-helices in chaperone activity, particularly in (H) samples suggesting a reversible conformational transition from helices to sheets.
This enabled us to formulate a functional model for binding of Acr to insulin B chains which incorporated 4 types of secondary structure molecules.
This might be a useful tool for analyzing in vitro preparations of recombinant Acr and build more consensuses on the structure-activity relationship especially in terms of oligomeric ratios.
نمط استشهاد جمعية علماء النفس الأمريكية (APA)
Krishnan, Gautam& Roy, Utpal. 2019. A New Functional Model for Prediction of Chaperone Activity of the Recombinant M. tb Acr (α-Crystallin) Using Insulin as Substrate. Canadian Journal of Infectious Diseases and Medical Microbiology،Vol. 2019, no. 2019, pp.1-18.
https://search.emarefa.net/detail/BIM-1130025
نمط استشهاد الجمعية الأمريكية للغات الحديثة (MLA)
Krishnan, Gautam& Roy, Utpal. A New Functional Model for Prediction of Chaperone Activity of the Recombinant M. tb Acr (α-Crystallin) Using Insulin as Substrate. Canadian Journal of Infectious Diseases and Medical Microbiology No. 2019 (2019), pp.1-18.
https://search.emarefa.net/detail/BIM-1130025
نمط استشهاد الجمعية الطبية الأمريكية (AMA)
Krishnan, Gautam& Roy, Utpal. A New Functional Model for Prediction of Chaperone Activity of the Recombinant M. tb Acr (α-Crystallin) Using Insulin as Substrate. Canadian Journal of Infectious Diseases and Medical Microbiology. 2019. Vol. 2019, no. 2019, pp.1-18.
https://search.emarefa.net/detail/BIM-1130025
نوع البيانات
مقالات
لغة النص
الإنجليزية
الملاحظات
Includes bibliographical references
رقم السجل
BIM-1130025
قاعدة معامل التأثير والاستشهادات المرجعية العربي "ارسيف Arcif"
أضخم قاعدة بيانات عربية للاستشهادات المرجعية للمجلات العلمية المحكمة الصادرة في العالم العربي
تقوم هذه الخدمة بالتحقق من التشابه أو الانتحال في الأبحاث والمقالات العلمية والأطروحات الجامعية والكتب والأبحاث باللغة العربية، وتحديد درجة التشابه أو أصالة الأعمال البحثية وحماية ملكيتها الفكرية. تعرف اكثر