Separation of Neurospora crassa myo-inositol-1-phosphate synthase from glucose-6-phosphate dehydrogenase by affinity chromatography.

J. Aradi, A. Zsindely, A. Kiss, M. Szabolcs, M. Schablik

Research output: Contribution to journalArticle

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Abstract

The purification of Neurospora crassa myo-inositol-1-phosphate synthase (EC 5.5.1.4) was studied by affinity chromatography using the substrate (glucose-6-phosphate), the inhibitor (pyrophosphate), the coenzyme (NAD+) and the coenzyme analogues (5'AMP and Cibacron Blue F3G-A) of the enzyme as adsorbents attached to agarose gel. Myo-inositol-1-phosphate synthase could be separated completely from the contaminating substance, glucose-6-phosphate dehydrogenase (EC 1.1.1.49), on Blue Sepharose CL-6B and on pyrophosphate-Sepharose. The purified enzyme had a specific activity of 16 400 U/mg. The sodium dodecyl sulfate/polyacrylamide gel electrophoresis of the 60 micrograms of this purified enzyme gave a homogenous band. The enzyme was found to be composed of four identical subunits having a molecular weight of 65 000.

Original languageEnglish
Pages (from-to)137-151
Number of pages15
JournalPreparative Biochemistry
Volume12
Issue number2
Publication statusPublished - 1982

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Myo-Inositol-1-Phosphate Synthase
Affinity chromatography
Neurospora crassa
Glucosephosphate Dehydrogenase
Affinity Chromatography
Cibacron Blue F 3GA
Coenzymes
Enzymes
Sepharose
Glucose-6-Phosphate
Adenosine Monophosphate
Electrophoresis
Sodium Dodecyl Sulfate
NAD
Adsorbents
Purification
Polyacrylamide Gel Electrophoresis
Molecular Weight
Gels
Molecular weight

ASJC Scopus subject areas

  • Biochemistry
  • Genetics

Cite this

Separation of Neurospora crassa myo-inositol-1-phosphate synthase from glucose-6-phosphate dehydrogenase by affinity chromatography. / Aradi, J.; Zsindely, A.; Kiss, A.; Szabolcs, M.; Schablik, M.

In: Preparative Biochemistry, Vol. 12, No. 2, 1982, p. 137-151.

Research output: Contribution to journalArticle

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