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[健康献血者和肾病患者血清蛋白组分的荧光分光光度分析]

[Spectrofluorometric analysis of protein fractions of blood serum of health donors and patients with kidney diseases].

作者信息

Parkhomenko T V, Klytsenko O A, Shavlovskiĭ M M, Poletaev A I, Kuznetsova I M, Turoverov K K

机构信息

St. Petersburg Pavlov Medical University.

出版信息

Tsitologiia. 1998;40(8-9):786-91.

PMID:9821250
Abstract

A comparison was made of ultra-violet fluorescence characteristics of albumin enriched protein fractions of serum blood of 35 glomerulonephritis (GN) patients, 30 uremic haemodialysis patients, and 40 donors. It has been found that fluorescence spectra of the albumin enriched fractions of serum from GN patients are blue shifted as compared with those of donors' serum preparations. At the same time fluorescence spectra of the albumin enriched fractions of donors' serum are similar, while those of GN patients vary significantly. The ratio of fluorescence intensities at 320 and 365 nm (A = I320/I365), characterizing the spectrum position is 1.27 +/- 0.05, for protein preparations of donors, while that for GN patients varies within the limits of 1.3-2.1. Fluorescence spectra of protein fractions from blood serum of uremic patients are red shifted (A = 0.77-1.29) in comparison with those of donors' blood. Chromatographic investigations show that protein preparations of GN patients' blood contain monomers and dimers of albumin, that can be divided according to their molecular masses or hydrophobic properties of the surface. Joint chromatographic and spectral analysis allowed to distinguish up to six albumin enriched fractions. A protein fraction with blue fluorescence spectrum was obtained by gel-filtration and ion exchange chromatography. A higher concentration of this fraction determines a high value of parameter A, that is typical for protein preparations of GN patients' blood. Amino acid sequence shows that one component of this fraction is beta-haptoglobin.

摘要

对35例肾小球肾炎(GN)患者、30例尿毒症血液透析患者和40例献血者血清中富含白蛋白的蛋白质组分的紫外荧光特性进行了比较。结果发现,与献血者血清制剂相比,GN患者血清中富含白蛋白的组分的荧光光谱发生了蓝移。同时,献血者血清中富含白蛋白的组分的荧光光谱相似,而GN患者的则有显著差异。表征光谱位置的320和365nm处荧光强度之比(A = I320/I365),献血者蛋白质制剂为1.27±0.05,而GN患者的则在1.3 - 2.1范围内变化。与献血者血液相比,尿毒症患者血清中蛋白质组分的荧光光谱发生了红移(A = 0.77 - 1.29)。色谱研究表明,GN患者血液中的蛋白质制剂含有白蛋白的单体和二聚体,可根据其分子量或表面的疏水性质进行区分。联合色谱和光谱分析可区分出多达六个富含白蛋白的组分。通过凝胶过滤和离子交换色谱获得了具有蓝色荧光光谱的蛋白质组分。该组分的较高浓度决定了参数A的高值,这是GN患者血液蛋白质制剂的典型特征。氨基酸序列表明该组分的一个成分是β-触珠蛋白。

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