Mass spectrometric quantification of asparagine synthetase in circulating leukemia cells from acute lymphoblastic leukemia patients
Mass spectrometric quantification of asparagine synthetase in circulating leukemia cells from acute lymphoblastic leukemia patients
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DOI：
10.1016/j.jprot.2007.11.009
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发表时间：
2008-04-30
影响因子：
3.3
通讯作者：
Conrads, Thomas P.
中科院分区：
文献类型：
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作者：
Abbatiello, Susan E.;Pan, Yuan-Xiang;Conrads, Thomas P.
天冬酰胺酶耐药的急性淋巴细胞白血病(ALL)在转化的细胞系中的出现与天冬酰胺合成酶(ASNS)的表达增加有关。然而，最近使用基于信使核糖核酸的分析方法进行的测量引起了人们对ASNS蛋白在赋予白血病细胞耐药性的细胞机制中的重要性的怀疑。因此，需要进行旨在确定人类白血病中ASNS蛋白浓度的研究来解决这一问题。提出了一种基于质谱仪(MS)的复杂样品混合物中ASNS蛋白质浓度的直接定量方法。这种检测方法能够区分样本和转化的细胞株，这些细胞株在很大的动态浓度范围内表达ASN。重要的是，这种方法直接检测ASNS蛋白，这种功能实体可能是合成足够的天冬酰胺，使白血病细胞对天冬酰胺酶治疗产生抗药性。我们还报告了这种MS方法的成功使用，它的检测和定量下限分别为30个和100个attomoles，首次直接测量了四个患者BLAST样本中ASNS蛋白的浓度。(C)2007 Elsevier B.V.保留所有权利。
The appearance of asparaginase-resistant acute lymphoblastic leukemia (ALL) in transformed cell lines has been correlated with increased expression of asparagine synthetase (ASNS). Recent measurements using mRNA-based assays have raised doubts, however, as to the importance of ASNS protein in the cellular mechanisms that confer drug resistance upon the leukemic cells. Studies aimed at determining the concentration of ASNS protein in human leukemias are therefore needed to resolve this issue. A mass spectrometry (MS)-based procedure is presented for the direct quantification of ASNS protein concentration in complex sample mixtures. This assay is able to distinguish samples from transformed cell lines that express ASNS over a wide dynamic range of concentration. Importantly, this method directly detects ASNS protein, the functional entity that may be synthesizing sufficient asparagine to render leukemia cells resistant to asparaginase-treatment. We also report the successful use of this MS method, which has lower limits of detection and quantification of 30 and 100 attomoles, respectively, for the first direct measurements of ASNS protein concentrations in four patient blast samples. (C) 2007 Elsevier B.V. All rights reserved.