---
_id: '12294'
abstract:
- lang: eng
  text: Brassicaceae are blessed with specialized metabolites called glucosinolates
    (GSLs), which along with their degradation products, are beneficial in agriculture
    and human health. To date, more than 130 GSL structures have been identified,
    mostly derived from the amino acid methionine. The biosynthesis of methionine-derived
    aliphatic GSLs starts with a side-chain elongation step involving a recursive
    three-step cyclic process that incorporates a new methylene group into the 2-oxo
    acid to form a series of elongated 2-oxo acids. Methylthioalkylmalate synthase
    (MAMS) catalyzes the first committed step in the side-chain elongation of methionine-derived
    GSLs. The substrate specificity of MAMS with different 2-oxo acids determines
    whether reaction products of a given cycle enter for an additional round of chain
    elongation or enter into core GSLs structure formation. Multiple MAMS encoding
    genes are present in the Brassicaceae species and are known to play a central
    role in shaping the diverse profile of aliphatic GSLs. We recently established
    a highly sensitive LC-MS/MS-based methodology that quantifies the MAMS activity
    by estimating the amount of the next intermediate of the pathway, the 2-malate
    derivatives. Overall, this chapter describes the protocol for the expression,
    purification, and steady-state kinetic analysis of the recombinant MAMS protein.
alternative_title:
- Methods in Enzymology
article_processing_charge: No
author:
- first_name: Roshan
  full_name: Kumar, Roshan
  id: 50e95140-3016-11ec-9e1a-881b648a1367
  last_name: Kumar
  orcid: 0000-0002-0346-9125
- first_name: Michael
  full_name: Reichelt, Michael
  last_name: Reichelt
- first_name: Naveen C.
  full_name: Bisht, Naveen C.
  last_name: Bisht
citation:
  ama: 'Kumar R, Reichelt M, Bisht NC. An LC-MS/MS Assay for Enzymatic Characterization
    of Methylthioalkylmalate Synthase (MAMS) Involved in Glucosinolate Biosynthesis.
    In: Jez J, ed. <i>Biochemical Pathways and Environmental Responses in Plants:
    Part A</i>. Vol 676. Elsevier; 2022:49-69. doi:<a href="https://doi.org/10.1016/bs.mie.2022.07.019">10.1016/bs.mie.2022.07.019</a>'
  apa: 'Kumar, R., Reichelt, M., &#38; Bisht, N. C. (2022). An LC-MS/MS Assay for
    Enzymatic Characterization of Methylthioalkylmalate Synthase (MAMS) Involved in
    Glucosinolate Biosynthesis. In J. Jez (Ed.), <i>Biochemical Pathways and Environmental
    Responses in Plants: Part A</i> (Vol. 676, pp. 49–69). Elsevier. <a href="https://doi.org/10.1016/bs.mie.2022.07.019">https://doi.org/10.1016/bs.mie.2022.07.019</a>'
  chicago: 'Kumar, Roshan, Michael Reichelt, and Naveen C. Bisht. “An LC-MS/MS Assay
    for Enzymatic Characterization of Methylthioalkylmalate Synthase (MAMS) Involved
    in Glucosinolate Biosynthesis.” In <i>Biochemical Pathways and Environmental Responses
    in Plants: Part A</i>, edited by Joseph Jez, 676:49–69. Elsevier, 2022. <a href="https://doi.org/10.1016/bs.mie.2022.07.019">https://doi.org/10.1016/bs.mie.2022.07.019</a>.'
  ieee: 'R. Kumar, M. Reichelt, and N. C. Bisht, “An LC-MS/MS Assay for Enzymatic
    Characterization of Methylthioalkylmalate Synthase (MAMS) Involved in Glucosinolate
    Biosynthesis,” in <i>Biochemical Pathways and Environmental Responses in Plants:
    Part A</i>, vol. 676, J. Jez, Ed. Elsevier, 2022, pp. 49–69.'
  ista: 'Kumar R, Reichelt M, Bisht NC. 2022.An LC-MS/MS Assay for Enzymatic Characterization
    of Methylthioalkylmalate Synthase (MAMS) Involved in Glucosinolate Biosynthesis.
    In: Biochemical Pathways and Environmental Responses in Plants: Part A. Methods
    in Enzymology, vol. 676, 49–69.'
  mla: 'Kumar, Roshan, et al. “An LC-MS/MS Assay for Enzymatic Characterization of
    Methylthioalkylmalate Synthase (MAMS) Involved in Glucosinolate Biosynthesis.”
    <i>Biochemical Pathways and Environmental Responses in Plants: Part A</i>, edited
    by Joseph Jez, vol. 676, Elsevier, 2022, pp. 49–69, doi:<a href="https://doi.org/10.1016/bs.mie.2022.07.019">10.1016/bs.mie.2022.07.019</a>.'
  short: 'R. Kumar, M. Reichelt, N.C. Bisht, in:, J. Jez (Ed.), Biochemical Pathways
    and Environmental Responses in Plants: Part A, Elsevier, 2022, pp. 49–69.'
date_created: 2023-01-16T10:05:13Z
date_published: 2022-10-21T00:00:00Z
date_updated: 2023-12-12T08:11:08Z
day: '21'
doi: 10.1016/bs.mie.2022.07.019
editor:
- first_name: Joseph
  full_name: Jez, Joseph
  last_name: Jez
external_id:
  pmid:
  - '36280361'
intvolume: '       676'
language:
- iso: eng
month: '10'
oa_version: None
page: 49-69
pmid: 1
publication: 'Biochemical Pathways and Environmental Responses in Plants: Part A'
publication_identifier:
  isbn:
  - '9780323955614'
  issn:
  - 0076-6879
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: An LC-MS/MS Assay for Enzymatic Characterization of Methylthioalkylmalate Synthase
  (MAMS) Involved in Glucosinolate Biosynthesis
type: book_chapter
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 676
year: '2022'
...