Ferreyra Raúl G, Burgardt Noelia I, Milikowski Daniel, Melen Gustavo, Kornblihtt Alberto R, Dell' Angelica Esteban C, Santomé José A, Ermácora Mario R
Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Bernal, Argentina.
Arch Biochem Biophys. 2006 Sep 15;453(2):197-206. doi: 10.1016/j.abb.2006.06.024. Epub 2006 Jul 21.
The 14-kDa sterol carrier protein 2 (SCP2) domain is present in Eukaria, Bacteria and Archaea, and has been implicated in the transport and metabolism of lipids. We report the cloning, expression, purification and physicochemical characterization of a SCP2 from the yeast Yarrowia lipolytica (YLSCP2). Analytical size-exclusion chromatography, circular dichroism and fluorescence spectra, indicate that recombinant YLSCP2 is a well-folded monomer. Thermal unfolding experiments show that SCP2 maximal stability is at pH 7.0-9.0. YLSCP2 binds cis-parinaric acid and palmitoyl-CoA with KD values of 81+/-40 nM and 73+/-33 nM, respectively, sustaining for the first time the binding of fatty acids and their CoA esters to a nonanimal SCP2. The role of yeast SCP2 and other lipid binding proteins in transport, storage and peroxisomal oxidation of fatty acids is discussed.
14千道尔顿的固醇载体蛋白2（SCP2）结构域存在于真核生物、细菌和古细菌中，并且与脂质的运输和代谢有关。我们报道了来自解脂耶氏酵母的一种SCP2（YLSCP2）的克隆、表达、纯化及物理化学特性。分析型尺寸排阻色谱、圆二色光谱和荧光光谱表明，重组YLSCP2是一种折叠良好的单体。热变性实验表明，SCP2的最大稳定性处于pH 7.0 - 9.0。YLSCP2分别以81±40 nM和73±33 nM的解离常数（KD）结合顺式十八碳四烯酸和顺式-9-十六碳烯酸辅酶A，首次证实了脂肪酸及其辅酶A酯与非动物SCP2的结合。本文还讨论了酵母SCP2和其他脂质结合蛋白在脂肪酸运输、储存和过氧化物酶体氧化中的作用。
