Galland A V, Dory D, Pons L, Chopin C, Rabesona H, Guéant J L, Fleurence J
Laboratoire de Pathologie Cellulaire et Moléculaire en Nutrition, EP CNRS 0616, Faculté de Médecine, Vandoeuvre-lès-Nancy, France.
J Chromatogr B Biomed Sci Appl. 1998 Feb 27;706(1):63-71. doi: 10.1016/s0378-4347(97)00457-x.
Cod fish is one of the foods most frequently involved in allergy. Only the cod allergen Gad c I, a 12.3 kDa parvalbumin, has been purified and characterized. Recently, we have detected allergen bands which have not previously been described, in particular a 41 kDa protein, by Western-blot. In the present work, this protein has been purified from a crude cod extract by ammonium sulfate fractionation, hydroxyapatite chromatography and preparative electrophoresis; a single band with an Mr of 41 x 10(3) was found in silver-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amino acid composition and the isoelectric point of the protein were determined. The purified protein (p41) was shown to bind specifically to reaginic IgE from sera of cod-allergic individuals and to a monoclonal anti-parvalbumin which recognizes specifically the first calcium binding site of parvalbumins. p41 may therefore contain a calcium binding site corresponding to an IgE-epitope similar to that of Gad c I.
鳕鱼是最常引发过敏的食物之一。仅有鳕鱼过敏原Gad c I，一种12.3 kDa的小清蛋白，得到了纯化和鉴定。最近，我们通过蛋白质免疫印迹法检测到了此前未被描述过的过敏原条带，尤其是一种41 kDa的蛋白质。在本研究中，该蛋白质已通过硫酸铵分级分离、羟基磷灰石层析和制备电泳从鳕鱼粗提物中纯化出来；在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳的银染中发现了一条Mr为41×10³的单一条带。测定了该蛋白质的氨基酸组成和等电点。纯化后的蛋白质（p41）被证明能特异性结合来自鳕鱼过敏个体血清中的反应素型IgE以及一种特异性识别小清蛋白第一个钙结合位点的抗小清蛋白单克隆抗体。因此，p41可能含有一个与Gad c I类似的IgE表位相对应的钙结合位点。
