Haginaka J, Seyama C, Kanasugi N
Faculty of Pharmaceutical Sciences, Mukogawa Women's University, Nishinomiya, Japan.
Anal Chem. 1995 Aug 1;67(15):2539-47. doi: 10.1021/ac00111a008.
Commercial chicken ovomucoid (OMCHI) and OMCHI, isolated by precipitation of egg whites with organic solvents, both of which as crude products, were fractionated by anion- and cation-exchange chromatography. The obtained four fractions were characterized by reversed-phase chromatography, N-terminal sequencing, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry, determination of sugar contents, and trypsin-inhibitory activities. Three fractions were OMCHI variants differing in carbohydrate composition, especially in sialic acid content, and the other fraction was tentatively termed ovoglycoprotein (OGCHI). The OMCHI and OGCHI are different in physicochemical and biochemical properties: average molecular weight 26000-27700 for OMCHI variants and 29700 for OGCHI; N-terminal amino acid, Ala for OMCHI and Thr for OGCHI; and trypsin-inhibitory activity, positive for OMCHI and negative for OGCHI. These OMCHI variants and OGCHI were bound to aminopropyl silica gels to evaluate chiral recognition ability. OMCHI is reported to have chiral recognition ability (Miwa, T.; et al. Chem. Pharm. Bull. 1987, 35, 682-686). However, neither OMCHI variant had appreciable chiral recognition ability, while the OGCHI had excellent chiral recognition properties as compared to those of the OMCHI reported previously. This reveals that the chiral recognition ability of the OMCHI reported previously comes from the OGCHI, which is present in crude OMCHI as an impurity.
市售鸡卵类粘蛋白（OMCHI）以及通过用有机溶剂沉淀蛋清分离得到的OMCHI（二者均为粗制品），通过阴离子交换色谱和阳离子交换色谱进行分级分离。对得到的四个级分进行反相色谱、N端测序、基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱分析、糖含量测定以及胰蛋白酶抑制活性测定来表征。其中三个级分为OMCHI变体，其碳水化合物组成不同，尤其是唾液酸含量不同，另一个级分暂称为卵糖蛋白（OGCHI）。OMCHI和OGCHI在物理化学和生化性质方面存在差异：OMCHI变体的平均分子量为26000 - 27700，OGCHI为29700；N端氨基酸，OMCHI为丙氨酸，OGCHI为苏氨酸；胰蛋白酶抑制活性，OMCHI呈阳性，OGCHI呈阴性。将这些OMCHI变体和OGCHI结合到氨丙基硅胶上以评估手性识别能力。据报道OMCHI具有手性识别能力（Miwa, T.; 等人，《化学与药学通报》，1987年，35卷，682 - 686页）。然而，OMCHI变体均没有明显的手性识别能力，而与先前报道的OMCHI相比，OGCHI具有优异的手性识别特性。这表明先前报道的OMCHI的手性识别能力来自OGCHI，OGCHI作为杂质存在于粗制OMCHI中。
