Panati Kalpana, Narala Venkata Ramireddy, Narasimha Vydyanath R, Derangula Madhavi, Arva Tatireddigari Venkat R R, Yeguvapalli Suneetha
Department of Biotechnology, Government College for Men, Kadapa, AP 516 004, India.
Department of Biotechnology, Sri Venkateswara University, Tirupati, AP 517 502, India.
J Genet Eng Biotechnol. 2018 Dec;16(2):459-466. doi: 10.1016/j.jgeb.2018.06.007. Epub 2018 Jul 2.
Fibronectin type III domain containing 5 (FNDC5) is a transmembrane protein. Upon cleavage, it yields a peptide called irisin that is supposedly bind to an unknown receptor and facilitates browning of white adipose tissue (WAT). Increased levels of irisin are associated with increased levels of energy expenditure markers PGC-1α, UCP-1, besides abundance of beige adipocytes in WAT. Though varied sizes of irisin were reported in humans and rodents it is not yet clear about the actual size of the irisin produced physiologically. Hence, we cloned and expressed human irisin (32-143 aa of FNDC5) in based on the proposed cleavage site that yields 12.5 kDa peptide to study its antigenicity and other biological functions . We purified recombinant human irisin (rh-irisin) to 95% homogeneity with simple purification method with a yield of 25 mg/g wet cell pellet. rh-irisin has been detected by commercially available antibodies from different sources with similar antigenicity. Biological activity of the rh-irisin was confirmed by using 3T3-L1 pre-adipocyte differentiation by Oil red O staining. Further, rh-irisin treatment on pre-adipocytes showed increased expression of markers associated with energy expenditure. As it is involved in energy expenditure process, it could be considered as potential therapeutic option for various metabolic diseases.
含III型纤连蛋白结构域5（FNDC5）是一种跨膜蛋白。经切割后，它会产生一种名为鸢尾素的肽，该肽可能与一种未知受体结合，并促进白色脂肪组织（WAT）的褐色化。鸢尾素水平升高与能量消耗标志物PGC-1α、解偶联蛋白1（UCP-1）水平升高以及WAT中米色脂肪细胞数量增加有关。尽管在人类和啮齿动物中报道了不同大小的鸢尾素，但尚不清楚生理产生的鸢尾素的实际大小。因此，我们根据产生12.5 kDa肽的假定切割位点，在[具体表达系统未给出]中克隆并表达了人鸢尾素（FNDC5的32 - 143氨基酸），以研究其抗原性和其他生物学功能。我们通过简单的纯化方法将重组人鸢尾素（rh - 鸢尾素）纯化至95%的纯度，产率为25 mg/g湿细胞沉淀。rh - 鸢尾素已被来自不同来源的市售抗体检测到，具有相似的抗原性。通过油红O染色利用3T3 - L1前脂肪细胞分化证实了rh - 鸢尾素的生物活性。此外，对前脂肪细胞进行rh - 鸢尾素处理显示与能量消耗相关的标志物表达增加。由于它参与能量消耗过程，它可被视为各种代谢疾病的潜在治疗选择。
