Harrison Steven L, Housden Nicholas G, Bottomley Stephen P, Cossins Aimee J, Gore Michael G
Biomolecular Sciences Group, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK.
Protein Expr Purif. 2008 Mar;58(1):12-22. doi: 10.1016/j.pep.2007.11.007. Epub 2007 Nov 22.
The Ig-binding properties of protein L from Peptostreptococcus magnus and protein G from Streptococcus have been successfully combined through the construction of a novel hybrid protein, consisting of a single Ig-binding domain from each protein. The biophysical and biochemical properties of this construct have been characterized through equilibrium and pre-equilibrium fluorescence spectroscopy, circular dichroism, isothermal titration calorimetry, affinity chromatography, and conformational stability studies using a chemical denaturant in order to examine the structure and availability of ligand binding sites in each domain. These studies show that despite the small size of the protein (Mw=16.5 kDa) each domain behaves in an independent manner with respect to the binding characteristics of the same domain in isolation.
通过构建一种新型杂合蛋白，成功地将大消化链球菌蛋白L和链球菌蛋白G的Ig结合特性结合在一起，该杂合蛋白由每种蛋白的单个Ig结合结构域组成。已通过平衡和预平衡荧光光谱、圆二色性、等温滴定量热法、亲和色谱以及使用化学变性剂的构象稳定性研究对该构建体的生物物理和生化特性进行了表征，以检查每个结构域中配体结合位点的结构和可用性。这些研究表明，尽管该蛋白尺寸较小（分子量=16.5 kDa），但每个结构域在结合特性方面的表现与单独存在时相同结构域的表现独立。
