Rout Ashok K, Minda R, Peri D, Ramakrishnan V, Bhattacharjee S K, Rao B J, Chary K V R
Department of Chemical Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Colaba, Mumbai, 400005, India.
Biomol NMR Assign. 2010 Oct;4(2):171-4. doi: 10.1007/s12104-010-9239-4. Epub 2010 Jun 5.
The cDNA of UVI31+ was cloned from C. reinhardtii and expressed in E. coli from where the protein was purified to homogeneity. The purified protein exhibited beta-lactamase activity (Manuscript in preparation). However, UVI31+ has no homology with the known β-lactamases. In order to understand the structural basis of the ability of UVI31+ to hydrolyze β-lactam antibiotics, we in parallel, set out to structurally characterize it by NMR. Its β-lactamase activity in relation to the solution structure by NMR is likely to provoke deeper understanding of its mechanism and facilitate the rationalization of other functions of the protein, if any. In this endeavor, we report almost complete sequence-specific backbone (1)H, (13)C and (15)N NMR assignments of UVI31+.
UVI31+的cDNA从莱茵衣藻中克隆出来，并在大肠杆菌中表达，然后从大肠杆菌中纯化出均一的蛋白质。纯化后的蛋白质表现出β-内酰胺酶活性（手稿正在准备中）。然而，UVI31+与已知的β-内酰胺酶没有同源性。为了了解UVI31+水解β-内酰胺抗生素能力的结构基础，我们同时着手通过核磁共振（NMR）对其进行结构表征。其β-内酰胺酶活性与通过NMR得到的溶液结构之间的关系，可能会促使我们更深入地了解其作用机制，并有助于阐明该蛋白质的其他功能（如果有的话）。在这项工作中，我们报告了UVI31+几乎完整的序列特异性主链（1）H、（13）C和（15）N NMR归属。
