Okano T, Masuda S, Ishimine M, Murai J, Yamamoto Y, Kobayashi T
J Nutr Sci Vitaminol (Tokyo). 1984 Apr;30(2):91-100. doi: 10.3177/jnsv.30.91.
The binding proteins for 25-hydroxyvitamin D3(25-OH-D3) in rat lymph and plasma were purified to apparent homogeneity as determined by sodium dodecyl sulfate disc gel electrophoresis. The purification procedures included the following techniques: gel filtration on Sephadex G-100 and G-200; affinity chromatography on Blue Sepharose CL-6B; ion-exchange chromatography on DEAE-Sepharose CL-6B; chromatofocussing on a Mono P column. Both proteins from lymph and plasma were eluted with the same retention time from an isoelectric column at a pH of approximately 4.53 and showed nearly identical data on the analysis of amino acid composition. When specific anti-lymph 25-OH-D3-binding protein antiserum was prepared in a rabbit, and Ouchterlony immunodiffusion was performed, the same precipitate line was observed on both lymph and plasma binding proteins. These results strongly suggest that a common transport protein exists in both rat lymph and plasma and circulates in the blood-lymph system.
通过十二烷基硫酸钠圆盘凝胶电泳测定，大鼠淋巴液和血浆中25-羟基维生素D3（25-OH-D3）的结合蛋白被纯化至表观均一性。纯化步骤包括以下技术：在Sephadex G - 100和G - 200上进行凝胶过滤；在Blue Sepharose CL - 6B上进行亲和层析；在DEAE - Sepharose CL - 6B上进行离子交换层析；在Mono P柱上进行色谱聚焦。淋巴液和血浆中的两种蛋白在pH约为4.53的等电柱上以相同的保留时间洗脱，并且在氨基酸组成分析中显示出几乎相同的数据。当在兔体内制备特异性抗淋巴25-OH-D3结合蛋白抗血清并进行双向免疫扩散时，在淋巴液和血浆结合蛋白上观察到相同的沉淀线。这些结果强烈表明，大鼠淋巴液和血浆中存在一种共同的转运蛋白，并在血液-淋巴系统中循环。
