Isolation and characterization of 2 -microglobulin from urine of kidney transplant and multiple myeloma patients.
Isolation and characterization of 2 -microglobulin from urine of kidney transplant and multiple myeloma patients.
复制标题
肾移植和多发性骨髓瘤患者尿液中 2-微球蛋白的分离和表征。
DOI：
10.1016/0003-9861(72)90409-2
复制
发表时间：
1972
影响因子：
3.9
通讯作者：
K. Kithier
中科院分区：
文献类型：
--
作者：
J. Čejka;K. Kithier
@-微球蛋白是一种低分子量的正常人类蛋白，首次从威尔逊氏病和慢性镉中毒患者的尿液中分离出来(1)，这些患者已知与肾小管功能受损和尿液中相对大量的低分子量蛋白排泄有关。通过免疫扩散和放射免疫测定，发现-微球蛋白在正常人血清、尿液和脑脊液中以微量存在（l-3）。这种蛋白的生理功能尚不清楚。然而，最近Smithies和Poulik(4)发现µglobulin的氨基酸序列与免疫球蛋白G的恒定区相似，这表明这两种蛋白之间存在联系。最近，上述作者发现的同源性已被整个p -微球蛋白分子的氨基酸序列所证实(5)。在我们对肾移植患者尿液中低分子蛋白排泄的研究过程中(6)，大量的&微球蛋白（在24例患者中高达100 mg）。在大多数患者的尿液中发现了Hr标本)。此外，在一些多发性骨髓瘤患者中，已发现蝇微球蛋白和其他低分子量蛋白质的含量增加。在目前的交流中，我们概述了一种用肾移植和骨髓瘤患者的尿液作为起始材料纯化微球蛋白的方法，对纯化蛋白进行了理化表征，并将我们的结果与Berggard和Bearn的结果进行了比较(1)。也有人试图找到纯化的&微球蛋白和免疫球蛋白G的多肽链及其片段之间的抗原关系。&-从6例肾移植患者和2例多发性骨髓瘤患者的尿液中分离纯化微球蛋白。24小时尿液标本经蒸馏水透析过夜，并通过UM-2型Diaflo膜进行超滤；保留液在洗脱缓冲液中透析后，将浓缩的尿蛋白涂在Bio-Gel P-100柱上（图1）。&-微球蛋白以组分5和至少6洗脱
@-Microglobulin, a normal human protein of low molecular weight, was first isolated (1) from urine of patients with Wilson’s disease and chronic cadmium poisoning, conditions known to be associated with impaired tubular function and excretion of relatively large amounts of low molecular-weight proteins in urine. By immunodiffusion and radioimmunoassay,, &-microglobulin was shown to be present in minute quantities in normal human serum, urine and cerebrospinal fluid (l-3). The physiological funct, ion of this protein is unknown. However, the recent finding by Smithies and Poulik(4) of similarities in the amino acid sequences of µglobulin and the constant region of immunoglobulin G suggested a relation between these two proteins. More recently, the homologies discovered by the above authors have been confirmed by the amino acid sequence of the entire p,-microglobulin molecule (5).In the course of our study of low molecularweight proteins excreted in urine of kidney transplant patients(6), large quantities of &-microglobulin (as much as 100 mg in 24. hr specimens) have been found in urine of most of these patients. Also, in some multiple myeloma patients increased amounts of fly-microglobulin, together with other low molecular-weight proteins, have been found. In the present communication we have outlined a method of purification of µglobulin using urines from kidney transplant and myeloma patients as a starting material, characterized physicochemically the purified protein, and compared our results with those of Berggard and Bearn (1). An attempt has also been made to find an antigenie relationship between the purified &-microglobulin and polypeptide chains of immunoglobulin G and its fragments. &-Microglobulin was purified separately from urines of six kidney transplant patients and two multiple myeloma patients. Twenty-four-hour urine specimens were dialyzed overnight against distilled water and ultrafiltrated through Diaflo, type UM-2 membranes; after dialysis of the retentate against the elution buffer, the concentrated urinary proteins were applied on a Bio-Gel P-100 column (Fig. 1). &-Microglobulin was eluted in fraction 5, together with at least six