Aron J, Baldwin D A, Marques H M, Pratt J M, Adams P A
J Inorg Biochem. 1986 Aug;27(4):227-43. doi: 10.1016/0162-0134(86)80064-2.
The heme-octapeptide from cytochrome c, Microperoxidase-8 (MP-8), was prepared by peptic and tryptic digestion of horse heart cytochrome c and purified by gel permeation chromatography in about 50% yield. Conditions for the identification of MP-8 by TLC and analysis by HPLC are described. Study of the concentration-dependence of the absorption spectrum showed that at concentrations of less than or equal to 2.5 X 10(-5) M in aqueous solution at pH 7, 25 degrees C and mu = 0.1, MP-8 exists as an equilibrium mixture of monomers and dimers with KD = 1.17 +/- 0.02 X 10(5) M-1, decreasing to 1.21 +/- 0.02 X 10(4) M-1 and 2.16 +/- 0.21 X 10(3) M-1 in 20% and 50% (v/v) methanol:water mixtures, respectively. Comparison of the Soret region spectrum of monomeric MP-8 with other hemoproteins suggests that it is six-coordinate in aqueous solution with water and His as axial ligands.
细胞色素c的血红素八肽，即微过氧化物酶-8（MP-8），是通过对马心细胞色素c进行胃蛋白酶和胰蛋白酶消化制备而成，并通过凝胶渗透色谱法进行纯化，产率约为50%。文中描述了通过薄层色谱法鉴定MP-8以及通过高效液相色谱法进行分析的条件。对吸收光谱浓度依赖性的研究表明，在pH 7、25℃且μ = 0.1的水溶液中，当MP-8浓度小于或等于2.5×10⁻⁵ M时，它以单体和二聚体的平衡混合物形式存在，KD = 1.17 ± 0.02×10⁵ M⁻¹，在20%（v/v）甲醇：水混合物和50%（v/v）甲醇：水混合物中分别降至1.21 ± 0.02×10⁴ M⁻¹和2.16 ± 0.21×10³ M⁻¹。将单体MP-8的Soret区光谱与其他血红蛋白进行比较表明，它在水溶液中以水和组氨酸作为轴向配体，呈六配位状态。
