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A new technique to track protein maturation

A new technique to track protein maturation

In an article entitled, “Atomic-resolution monitoring of protein maturation in live human cells by NMR,” published on March 3, 2013 issue of Nature Chemical Biology (dOI:10.1038/nCHeMBIO.1202), the lead author Lucia Banci with six other associates affiliated to the  University of Florence, Italy & the University of Oxford, UK have described a new technique to track proteins at atomic resolution in live cells.   Applying this technique,  investigators have been able to observe how proteins fold and mature into functional forms. They also applied nuclear magnetic resonance spectroscopy (NMR) directly in live human cells to depict the absolute post-translational maturation procedure of human superoxide dismutase 1 (SOD1). They followed zinc binding, homodimer formation and copper uptake at atomic resolution and discovered that copper chaperone for SOD1 oxidizes the SOD1 intra-subunit disulfide bond through both copper-dependent and copper-independent mechanisms. Their proposed approach represents a new strategy for structural investigation of endogenously expressed proteins in a physiological (cellular) environment. The method also showed that disulphide bonds can form even when SOD1 has not bound copper, a result that has not been seen using purified proteins. However, successful NMR analysis of endogenously expressed proteins in live mammalian cells requires efficient cDNA transfection, relatively high protein expression levels, and the application of different labeling strategies and the maintenance of cell integrity during measurements. In this study, researchers have addressed all of these aspects and followed the sequential physiological order of the events in the SOD1 post-translational modification process to obtain information that cannot be retrieved in vitro. This strategy is potentially applicable for many other protein targets  thus opening the way for a broad range study of in-cell protein structures  at the molecular level. [Summarized by a graduate student, Samsad Razzaque, a graduate student at Plantbiotechlab, DU.]

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