Analysis of Sinapoylmalate in the Arabidopsis thaliana Leaf by Using the nano|Filter Vial®: Sinapoyl Malate is a major UV protectant in Arabidopsis thaliana

Data provided by Jing-Ke Weng, Ph.D.
Member, Whitehead Institute for Biomedical Research
Assistant Professor of Biology, Massachusetts Institute of Technology

Introduction

Plants contain rapidly evolving specialized metabolic system, and presumably encounter destabilized evolutionary intermediates along their mutational trajectories. Have plants evolved unique molecular mechanisms that assist folding of those destabilized proteins and/or mitigate proteotoxicity arising from protein misfolding? We use the model plant Arabidopsis thaliana to examine the in vivo function and behavior of mutant enzymes that exhibit broadened product promiscuity and/or decreased folding stability in vitro. We attempt to identify genetic components involved in cellular mechanisms that assist folding or alter product profile of these mutant enzymes. We designed a simple sample preparation method for the analysis of Sinapoyl Malate by LC-MS to study its mechanism in UV protection of the Arabidopsis thaliana.

Arabidopsis Leaf

Method

  • Grind Leaf tissue under liquid nitrogen.
  • Extract with 80% MeOH (1mL MeOH to 200mg fresh weight leaves).
  • Centrifuge.
  • Filter with 0.2um PTFE nano|Filter Vial™.
  • Analyze by UHPLC - Orbitrap Mass Spec.

Equipment

HPLC Column: Phenomenex Kinetex 2.6µm C18 150x3.0

Mobile Phase

Solvent A: H2O + 0.1% Formic Acid

Solvent B: Acetonitrile + 0.1% Formic Acid

Gradient

time%A%B
2min955%
40min2080%
40.1min595%
44min595%
44.1min955%
48min955%
Arabidopsis Leaf elements

Title

Analysis of Sinapoylmalate in the Arabidopsis thaliana Leaf by Using the nano|Filter Vial®: Sinapoyl Malate is a major UV protectant in Arabidopsis thaliana

Description

We use the model plant Arabidopsis thaliana to examine the in vivo function and behavior of mutant enzymes that exhibit broadened product promiscuity and/or decreased folding stability in vitro. We attempt to identify genetic components involved in cellular mechanisms that assist folding or alter product profile of these mutant enzymes.

PDF Application

Product Used in Application

nano|Filter Vial® - PTFE 0.45µm

Non-Split Septum, Blue Cap

pn#15540-200

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