کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8359233 | 1542289 | 2018 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Heterologous expression of the human polybromo-1 protein in the methylotrophic yeast Pichia pastoris
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کلمات کلیدی
AExBAF180PBRM1HmgPRRBRDNHEJBromodomain - برومودومینBah - بهاRecombinant protein expression - بیان پروتئین نوترکیبPost-replication repair - تعمیر پس از تکرارGAP - شکافnon-homologous end-joining - عدم پیوستن به همولوگPichia pastoris - پیکیا پاستوریسanion exchange chromatography - کروماتوگرافی تبادل آنیونhigh mobility group - گروه تحرک بالاglyceraldehyde-3-phosphate dehydrogenase - گلیسرالیدید-3-فسفات دهیدروژناز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Heterologous expression of the human polybromo-1 protein in the methylotrophic yeast Pichia pastoris Heterologous expression of the human polybromo-1 protein in the methylotrophic yeast Pichia pastoris](/preview/png/8359233.png)
چکیده انگلیسی
The human polybromo-1 protein (BAF180) is a known driver mutation in clear cell renal cell carcinoma, where it is mutated in approximately 40% of cases. BAF180 is the chromatin-targeting subunit of the PBAF complex. BAF180 has six bromodomains, two BAH domains, and one HMG box. Bromodomains are known to recognize acetylated-lysines on histones and play a role in nucleosome recognition. BAH domains are required for ubiquitination of PCNA, a key regulator of DNA damage. The putative HMG box, if functional, may be involved in DNA-binding. While the binding specificities of individual bromodomains have been studied by our lab and others, the results have failed to reach a consensus. The acetyl-histone binding features of the full-length protein is unknown and is the motivation for this work. The hypothetical HMG and BAH domains have not been studied and the actual function of these regions is currently unknown. Thus, the precise interactions of this large and complex protein are not well-studied. Advances in understanding this large protein have been hindered by the inability to express and purify recombinant full-length BAF180 protein. Currently, only phenomenological studies using BAF180 expressed in mammalian cells have been conducted. Here, we report the successful expression, purification of full-length biologically active BAF180 protein using the GAP promoter in the heterologous host Pichia pastoris. The ability to express full-length and mutated BAF180 will allow for biophysical binding studies. Knowledge of the binding interactions is critical for us to understand the role of BAF180 in cancer development and its progression.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 152, December 2018, Pages 23-30
Journal: Protein Expression and Purification - Volume 152, December 2018, Pages 23-30
نویسندگان
Sarah E. Hopson, Martin J. Thompson,