PMID: 11607197Jul 1, 1991Paper

In vitro assay of the chlorophyll biosynthetic enzyme Mg-chelatase: resolution of the activity into soluble and membrane-bound fractions

Proceedings of the National Academy of Sciences of the United States of America
C J Walker, J D Weinstein

Abstract

The first committed step in chlorophyll synthesis is the Mg-chelatase-catalyzed insertion of magnesium into protoporphyrin IX. Since iron insertion into protoporphyrin leads to heme formation, Mg-chelastase lies at the branch point of heme and chlorophyll synthesis in chloroplasts. Little is known about the enzymology or regulation of Mg-chelatase, as it has been assayed only in intact cucumber chloroplasts. In this report we describe an in vitro assay for Mg-chelatase. Mg-chelatase activity in intact pea chloroplasts was 3- to 4-fold higher than in cucumber chloroplasts. This activity survived chloroplast lysis and could be fractionated by centrifugation into supernatant and pellet components. Both of these fractions were required to reconstitute Mg-chelatase activity, and both were inactivated by boiling indicating that the enzyme is composed of soluble and membrane-bound protein(s). The product of the reaction was confirmed fluorometrically as the magnesium chelate of the porphyrin substrate. The specific activity of the reconstituted system was typically 1 nmol of Mg-deuteroporphyrin per h per mg of protein, and activity was linear for at least 60 min under our assay conditions. ATP and magnesium were required for Mg-chelat...Continue Reading

References

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Citations

Jan 1, 1994·Photosynthesis Research·J K HooberJ L Gabriel
Dec 1, 1992·Plant Molecular Biology·C KonczJ Schell
Mar 21, 2007·Photosynthesis Research·David W Bollivar
Mar 6, 1999·Current Opinion in Plant Biology·B Grimm
Jan 11, 2012·Journal of Experimental Botany·Olaf Czarnecki, Bernhard Grimm
Apr 15, 1996·European Journal of Biochemistry·S Reinbothe, C Reinbothe
Jan 15, 1996·European Journal of Biochemistry·R D WillowsD von Wettstein
Feb 15, 2001·Proceedings of the National Academy of Sciences of the United States of America·N MochizukiJ Chory
Dec 10, 2009·Molecular Plant·Enrico Peter, Bernhard Grimm
Mar 13, 2010·Structure·Joakim LundqvistSalam Al-Karadaghi
Jul 29, 2004·FEBS Letters·Annegret WildeBernhard Grimm
Jul 13, 2004·Plant Physiology and Biochemistry : PPB·Ulf OlssonMats Hansson
Jul 10, 2013·The Journal of Biological Chemistry·Joakim LundqvistMats Hansson
May 3, 2007·The Journal of Biological Chemistry·Akinori IkegamiTatsuru Masuda
Feb 12, 2008·The Journal of Biological Chemistry·Nickolche SirijovskiMats Hansson
Jul 17, 2009·The Journal of Biological Chemistry·Neil D AdhikariRobert M Larkin
Dec 12, 2019·G3 : Genes - Genomes - Genetics·Rajdeep S KhanguraGurmukh S Johal
Aug 9, 2003·The Plant Journal : for Cell and Molecular Biology·Elena YaronskayaBernhard Grimm
Jul 30, 2019·Theoretical and Experimental Plant Physiology·Pedro F N SouzaFabricio E L Carvalho

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