硫芥的生物学归宿：硫芥对人血红蛋白的体外烷基化作用。

Biological fate of sulphur mustard: in vitro alkylation of human haemoglobin by sulphur mustard.

Black R M, Harrison J M, Read R W

Chemical and Biological Defence Sector, Defence Evaluation and Research Agency, Porton Down, Salisbury, UK.

Xenobiotica. 1997 Jan;27(1):11-32. doi: 10.1080/004982597240730.

Abstract

Human blood was incubated in vitro with a 1:1 mixture of [35S,12C4]- and [13C4]-sulphur mustard. Alkylated globin, containing the 2-hydroxyethylthioethyl (HETE) moiety, was isolated from the blood incubate following lysis of the erythrocytes and acidification with HCl in isopropanol. 2. The alkylated globin was hydrolysed with Pronase E to give a digest containing alkylated amino acids and alkylated dipeptides. A number of these were partially purified by hplc and identified by gc-ms and lc-ms. 3. The alkylated globin was hydrolysed with trypsin to give a digest containing alkylated peptides. Ten of these were partially purified by hplc, tentatively identified by lc-electrospray mass spectrometry, and the sequences and sites of alkylation determined using lc-electrospray tandem mass spectrometry. 4. (2-Hydroxyethylthioethyl)glutathione was also shown to be present in the pronase and trypsin digests of alkylated globin. 5. N-terminal valine, on both the alpha and beta chains, and histidine residues were identified as the key sites of interaction for targeting as biological markers of sulphur mustard poisoning.

摘要

将人血与[35S,12C4]-硫芥和[13C4]-硫芥按1:1的混合物在体外孵育。红细胞裂解并用异丙醇中的盐酸酸化后，从血样孵育物中分离出含有2-羟乙硫基乙基（HETE）部分的烷基化珠蛋白。
用链霉蛋白酶E水解烷基化珠蛋白，得到含有烷基化氨基酸和烷基化二肽的消化物。其中一些通过高效液相色谱法进行部分纯化，并通过气相色谱-质谱联用仪和液相色谱-质谱联用仪进行鉴定。
用胰蛋白酶水解烷基化珠蛋白，得到含有烷基化肽段的消化物。其中十个通过高效液相色谱法进行部分纯化，通过液相色谱-电喷雾质谱法进行初步鉴定，并使用液相色谱-电喷雾串联质谱法确定烷基化的序列和位点。
还证明在烷基化珠蛋白的链霉蛋白酶和胰蛋白酶消化物中存在（2-羟乙硫基乙基）谷胱甘肽。
α链和β链上的N端缬氨酸以及组氨酸残基被确定为作为硫芥中毒生物标志物的靶向相互作用的关键位点。