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Related Experiment Video

Updated: Jul 24, 2026

Identification of Post-translational Modifications of Plant Protein Complexes
10:07

Identification of Post-translational Modifications of Plant Protein Complexes

Published on: February 22, 2014

Plant dihydroxyacetone phosphate reductases : purification, characterization, and localization.

T Kirsch1, D W Gerber, R U Byerrum

  • 1Department of Biochemistry, Michigan State University, East Lansing, Michigan 48824-1319.

Plant Physiology
|September 1, 1992
PubMed
Summary

Spinach leaves contain distinct dihydroxyacetone phosphate (DHAP) reductase enzymes in the cytosol, chloroplasts, and peroxisomes. These isoforms, while sharing some properties, exhibit unique characteristics and sensitivities, impacting plant metabolism.

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Area of Science:

  • Biochemistry
  • Plant Physiology
  • Enzymology

Background:

  • Dihydroxyacetone phosphate (DHAP) reductase plays a crucial role in metabolic pathways.
  • Understanding the different isoforms of DHAP reductase is essential for elucidating plant metabolic regulation.

Purpose of the Study:

  • To purify and characterize cytosolic DHAP reductase from spinach leaves.
  • To separate and partially purify a chloroplastic DHAP reductase isoform.
  • To isolate and identify a peroxisomal DHAP reductase isoform.

Main Methods:

  • Multi-step protein purification including anion-exchange, gel filtration, hydrophobic, and dye-ligand chromatography.
  • Antibody generation and cross-reactivity testing.
  • Enzyme activity assays and molecular weight determination.

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2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
08:23

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

Published on: August 6, 2018

Affinity Purification of Chloroplast Translocon Protein Complexes Using the TAP Tag
07:01

Affinity Purification of Chloroplast Translocon Protein Complexes Using the TAP Tag

Published on: November 1, 2018

Related Experiment Videos

Last Updated: Jul 24, 2026

Identification of Post-translational Modifications of Plant Protein Complexes
10:07

Identification of Post-translational Modifications of Plant Protein Complexes

Published on: February 22, 2014

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
08:23

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes

Published on: August 6, 2018

Affinity Purification of Chloroplast Translocon Protein Complexes Using the TAP Tag
07:01

Affinity Purification of Chloroplast Translocon Protein Complexes Using the TAP Tag

Published on: November 1, 2018

Main Results:

  • A cytosolic DHAP reductase was purified to homogeneity with specific activity of 500 mumol min(-1) mg(-1) and subunit molecular mass of 38 kD.
  • A distinct chloroplastic DHAP reductase isoform was separated, showing no cross-reactivity with antibodies against the cytosolic form.
  • A third DHAP reductase isoform was identified in peroxisomes, sensitive to antibodies against the cytosolic enzyme.

Conclusions:

  • Spinach leaves possess at least three distinct DHAP reductase isoforms located in the cytosol, chloroplasts, and peroxisomes.
  • These isoforms exhibit unique biochemical properties and immunological relationships.
  • DHAP reductase isoforms are specific for NADH and DHAP, with varying sensitivities to inhibitors and activators.