Analytical and preparative separation of the diastereomers of l-buthionine (SR)-sulfoximine, a potent inhibitor of glutathione biosynthesis

丁硫胺 谷胱甘肽 化学 立体化学 生物合成 生物化学
作者
Ernest B. Campbell,Michael L. Hayward,Owen W. Griffith
出处
期刊:Analytical Biochemistry [Elsevier BV]
卷期号:194 (2): 268-277 被引量:48
标识
DOI:10.1016/0003-2697(91)90229-m
摘要

Buthionine sulfoximine inhibits γ-glutamylcysteine synthetase, the enzyme catalyzing the first reaction of glutathione (GSH) biosynthesis. GSH synthesis is blocked in animals or cultured cells exposed to buthionine sulfoximine, and GSH is substantially depleted in cells or tissues with moderate to high rates of GSH utilization. Studies reported to date have used dl-buthionine (SR)-sulfoximine or l-buthionine (SR)-sulfoximine, mixtures of four and two isomers, respectively. The present report describes a chiral solvent HPLC procedure for the analytical separation of the diastereomers of l-buthionine (SR)-sulfoximine and the separation of those isomers from the unresolved diastereomers of l-buthionine (SR)-sulfoximine. l-Buthionine (R)-sulfoximine was isolated preparatively by repeated crystallization of l-buthionine (SR)-sulfoximine from water; l-buthionine (S)-sulfoximine was obtained by crystallization as the trifluoroacetate salt in ethanol/hexane mixtures. The absolute configuration, bond lengths and angles of l-buthionine (R)-sulfoximine were determined by X-ray diffraction. In vitro studies demonstrate that l-buthionine (R)-sulfoximine is a relatively weak inhibitor of rat kidney γ-glytamylcysteine synthetase; binding is competitive with l-glutamate. l-Buthionine (S)-sulfoximine is a tight-binding, mechanism-based inhibitor of the enzyme. Since l-buthionine sulfoximine is initially bound as a transition-state analogue, identification of the inhibitory diastereomer elucidates the steric relationships among ATP, glutamate, and cysteine within the active site. When administered to mice, l-buthionine (S)-sulfoximine (0.2 mmol/kg) was as effective as l-buthionine (SR)-sulfoximine (0.4 mmol/kg) in causing GSH depletion in liver, kidney, and pancreas. l-Buthionine (R)-sulfoximine (0.2 mmol/kg) did not cause significant GSH depletion in liver or pancreas. The l-(R)-diastereomer caused a modest GSH depletion in kidney that is tentatively attributed to interference with γ-glutamylcyst(e)ine transport.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
1秒前
3秒前
3秒前
在水一方应助迷路的忆之采纳,获得10
3秒前
Dead Cells完成签到,获得积分10
3秒前
着急的柔完成签到,获得积分10
3秒前
有魅力的小蜜蜂完成签到,获得积分10
4秒前
慕青应助lizhaonian采纳,获得10
4秒前
耍酷的傲霜完成签到,获得积分10
5秒前
善学以致用应助jjsun采纳,获得10
6秒前
EED应助文件撤销了驳回
6秒前
小蘑菇应助年轻的烨华采纳,获得10
7秒前
7秒前
zzz发布了新的文献求助10
8秒前
研友_Zzrx6Z完成签到,获得积分10
9秒前
9秒前
11秒前
11秒前
汉堡包应助NOTHING采纳,获得10
11秒前
SYLH应助科研通管家采纳,获得50
11秒前
quhayley应助科研通管家采纳,获得10
11秒前
爆米花应助科研通管家采纳,获得10
11秒前
坦率的匪应助科研通管家采纳,获得10
12秒前
田様应助科研通管家采纳,获得10
12秒前
SYLH应助科研通管家采纳,获得50
12秒前
orixero应助科研通管家采纳,获得10
12秒前
SYLH应助科研通管家采纳,获得50
12秒前
思源应助科研通管家采纳,获得10
12秒前
12秒前
隐形曼青应助科研通管家采纳,获得10
12秒前
czh应助科研通管家采纳,获得10
12秒前
赘婿应助科研通管家采纳,获得10
12秒前
华仔应助科研通管家采纳,获得10
12秒前
科研通AI2S应助科研通管家采纳,获得10
12秒前
13秒前
SYLH应助科研通管家采纳,获得50
13秒前
大模型应助科研通管家采纳,获得10
13秒前
13秒前
斯文败类应助Keyl采纳,获得10
13秒前
褪黑素应助科研通管家采纳,获得10
13秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
‘Unruly’ Children: Historical Fieldnotes and Learning Morality in a Taiwan Village (New Departures in Anthropology) 400
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3988732
求助须知:如何正确求助?哪些是违规求助? 3531027
关于积分的说明 11252281
捐赠科研通 3269732
什么是DOI,文献DOI怎么找? 1804764
邀请新用户注册赠送积分活动 881869
科研通“疑难数据库(出版商)”最低求助积分说明 809021