基于质谱法的光呼吸中间体定量分析

Quantification of Photorespiratory Intermediates by Mass Spectrometry-Based Approaches.

作者信息

Arrivault Stéphanie, Obata Toshihiro

机构信息

Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, Golm, 14476, Potsdam, Germany.

University of Nebraska-Lincoln, Department of Biochemistry, Center for Plant Science Innovation, E321 Beadle Center, 1901 Vine Street, Lincoln NE, 68588-0664, USA.

出版信息

Methods Mol Biol. 2017;1653:97-104. doi: 10.1007/978-1-4939-7225-8_7.

DOI:10.1007/978-1-4939-7225-8_7

PMID:28822128

Abstract

Photorespiration is an essential metabolic process in plants occurring via the oxygenase reaction of RuBisCO. In order to understand this process, it is essential to determine the amounts of intermediates involved. For this purpose we combined mass spectrometry-based approaches and the use of authentic standards for the quantification of photorespiratory intermediates. Here we describe protocols for the extraction and quantification of 2-phosphoglycolate (2PG) by LC-MS/MS and serine, glycine, glycolate, hydroxypyruvate, glyoxylate, and glycerate by GC-MS.

摘要

光呼吸是植物中通过核酮糖-1,5-二磷酸羧化酶/加氧酶（RuBisCO）的加氧酶反应发生的一个重要代谢过程。为了理解这个过程，确定所涉及的中间产物的量至关重要。为此，我们将基于质谱的方法与使用真实标准品来定量光呼吸中间产物相结合。在这里，我们描述了通过液相色谱-串联质谱法（LC-MS/MS）提取和定量2-磷酸乙醇酸（2PG）以及通过气相色谱-质谱法（GC-MS）提取和定量丝氨酸、甘氨酸、乙醇酸、羟基丙酮酸、乙醛酸和甘油酸的实验方案。