Expression and Purification of Active Recombinant Cathepsin C (Dipeptidyl Aminopeptidase I) of Kuruma Prawn Marsupenaeus japonicus in Insect Cells
المؤلفون المشاركون
Feng, Hai-Yang
Qiu, Gao-Feng
Yamano, Keisuke
المصدر
Journal of Biomedicine and Biotechnology
العدد
المجلد 2009، العدد 2009 (31 ديسمبر/كانون الأول 2009)، ص ص. 1-6، 6ص.
الناشر
Hindawi Publishing Corporation
تاريخ النشر
2009-08-18
دولة النشر
مصر
عدد الصفحات
6
التخصصات الرئيسية
الملخص EN
Cathepsin C (CTSC) is a lysosomal cysteine protease belonging to the papain superfamily.
Our previous study showed that CTSC precursor (zymogen) is localized exclusively in cortical rods (CRs) of mature oocyte in the kuruma prawn Marsupenaeus japonicus, suggesting that CTSC might have roles on regulating release and/or formation of a jelly layer.
In this study, enzymically active CTSC of the kuruma prawn was prepared by recombinant expression in the High Five insect cell line.
The recombinant enzyme with a polyhistidine tag at its C-terminus was considered to be initially secreted into the culture medium as an inactive form of zymogen, because Western blot with anti-CTSC antibody detected a 51 kDa protein corresponding to CTSC precursor.
After purification by affinity chromatography on nickel-iminodiacetic acid resin, the enzyme displayed three forms of 51, 31, and 30 kDa polypeptides.
All of the forms can be recognized by antiserum raised against C-terminal polyhistidine tag, indicating that the 31 and 30 kDa forms were generated from 51 kDa polypeptide by removal of a portion of the N-terminus of propeptide.
Following activation at pH 5.5 and 37∘C for 40 hours under native conditions, the recombinant CTSC (rCTSC) exhibited increased activity against the synthetic substrate Gly-Phe-β-naphthylamide and optimal pH at around 5.
The purified rCTSC will be useful for further characterization of its exact physiological role on CRs release and/or formation of a jelly layer in kuruma prawn.
نمط استشهاد جمعية علماء النفس الأمريكية (APA)
Qiu, Gao-Feng& Feng, Hai-Yang& Yamano, Keisuke. 2009. Expression and Purification of Active Recombinant Cathepsin C (Dipeptidyl Aminopeptidase I) of Kuruma Prawn Marsupenaeus japonicus in Insect Cells. Journal of Biomedicine and Biotechnology،Vol. 2009, no. 2009, pp.1-6.
https://search.emarefa.net/detail/BIM-495395
نمط استشهاد الجمعية الأمريكية للغات الحديثة (MLA)
Qiu, Gao-Feng…[et al.]. Expression and Purification of Active Recombinant Cathepsin C (Dipeptidyl Aminopeptidase I) of Kuruma Prawn Marsupenaeus japonicus in Insect Cells. Journal of Biomedicine and Biotechnology No. 2009 (2009), pp.1-6.
https://search.emarefa.net/detail/BIM-495395
نمط استشهاد الجمعية الطبية الأمريكية (AMA)
Qiu, Gao-Feng& Feng, Hai-Yang& Yamano, Keisuke. Expression and Purification of Active Recombinant Cathepsin C (Dipeptidyl Aminopeptidase I) of Kuruma Prawn Marsupenaeus japonicus in Insect Cells. Journal of Biomedicine and Biotechnology. 2009. Vol. 2009, no. 2009, pp.1-6.
https://search.emarefa.net/detail/BIM-495395
نوع البيانات
مقالات
لغة النص
الإنجليزية
الملاحظات
Includes bibliographical references
رقم السجل
BIM-495395
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