pmc logo image
Logo of plntphysJournal URL: http://www.plantphysiol.org
Journal List > Plant Physiol > v.92(3); Mar 1990

Formats:
Plant Physiol. 1990 March; 92(3): 710–717.
PMCID: PMC1062358
Copyright notice
Purification by Immunoadsorption and Immunochemical Properties of NADP-Dependent Malic Enzymes from Leaves of C3, C4, and Crassulacean Acid Metabolism Plants
Marion Fathi1 and Claus Schnarrenberger
Institute of Plant Physiology, Cell Biology and Microbiology, Free University of Berlin, Königin-Luise-Strasse 12-16a, D-1000 Berlin 33 (West), Federal Republic of Germany
1 Present address: Bundesgesundheitsamt, Thielallee 88-92, D-1000 Berlin 33 (West).
Small right arrow pointing to: This article has been cited by other articles in PMC.
Abstract
NADP:malic enzyme from corn (Zea mays L.) leaves was purified by conventional techniques to apparent homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antibodies raised against this protein in rabbits were purified, coupled covalently to protein A-Sepharose CL-4B, and used as an immunoaffinity resin to purify the NADP:malic enzymes of the C3 plants spinach (Spinacia oleracea L.) and wheat (Triticum aestivum L.), of the Crassulacean acid metabolism (CAM) plant Bryophyllum daigremontianum R. Hamed et Perr. de la Bathie and the C4 plants corn, sugarcane (Saccharum officinarum L.), and Portulaca grandiflora L. Such procedures yielded homogeneous protein preparations with a single protein band, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, except for P. grandiflora L. with two bands. The specific activities of the purified proteins ranged between 56 and 91 units (milligrams per protein). NADP:malic enzyme represented up to 1% of the total soluble protein in C4 plants, 0.5% in the CAM plant, and less than 0.01% in C3 plants. In immunotitration tests involving immunoprecipitation and immunoinhibition of activity by an antiserum against the corn leaf enzyme, the NADP:malic enzymes of corn and sugarcane showed virtually full identity of epitopes, while the NADP:malic enzymes of the C3 and CAM plants exhibited a cross-reaction of one-twentieth and one-fourth by these tests, respectively. The NADP:malic enzyme of P. grandiflora exhibited characteristics more closely related to the enzymes of C3 and CAM plants than to those of C4 plants.
Full text
Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.5M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.
 
icon of scanned page 710
710
icon of scanned page 711
711
icon of scanned page 712
712
icon of scanned page 713
713
icon of scanned page 714
714
icon of scanned page 715
715
icon of scanned page 716
716
icon of scanned page 717
717
 
Images in this article
Figure 1
Figure 1

Figure 1
on p.713
Figure 2
Figure 2

Figure 2
on p.715
Click on the image to see a larger version.
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
  • Asami S, Inoue K, Matsumoto K, Murachi A, Akazawa T. NADP-malic enzyme from maize leaf: purification and properties. Arch Biochem Biophys. 1979 May;194(2):503–510. [PubMed]
  • Bensadoun A, Weinstein D. Assay of proteins in the presence of interfering materials. Anal Biochem. 1976 Jan;70(1):241–250. [PubMed]
  • Böhme HJ, Kopperschläger G, Schulz J, Hofmann E. Affinity chromatography of phosphofructokinase using Cibacron blue F3G-A. J Chromatogr. 1972 Jun 28;69(1):209–214. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Brandon PC, van Boekel-Mol TN. Properties of purified malic enzyme in relation to crassulacean acid metabolism. Eur J Biochem. 1973 May;35(1):62–69. [PubMed]
  • Chapman KS, Hatch MD. Regulation of mitochondrial NAD-malic enzyme involved in C4 pathway photosynthesis. Arch Biochem Biophys. 1977 Nov;184(1):298–306. [PubMed]
  • Collins PD, Hague DR. Light-stimulated synthesis of NADP malic enzyme in leaves of maize. J Biol Chem. 1983 Mar 25;258(6):4012–4018. [PubMed]
  • Davies DD, Patil KD. Regulation of 'malic' enzyme of Solanum tuberosum by metabolites. Biochem J. 1974 Jan;137(1):45–53. [PubMed]
  • DAVIS BJ. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. [PubMed]
  • Gersten DM, Marchalonis JJ. A rapid, novel method for the solid-phase derivatization of IgG antibodies for immune-affinity chromatography. J Immunol Methods. 1978;24(3-4):305–309. [PubMed]
  • Hatch MD, Mau SL. Association of NADP- and NAD-linked malic enzyme acitivities in Zea mays: relation to C4 pathway photosynthesis. Arch Biochem Biophys. 1977 Mar;179(2):361–369. [PubMed]
  • Hatch MD, Mau SL, Kagawa T. Properties of leaf NAD malic enzyme from plants with C4 pathway photosynthesis. Arch Biochem Biophys. 1974 Nov;165(1):188–200. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • MARTIN RG, AMES BN. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed]
  • Mukerji SK, Ting IP. Malate dehydrogenase (decarboxylating) (NADP) isoenzymes of Opuntia stem tissue. Mitochondrial, chloroplast, and soluble forms. Biochim Biophys Acta. 1968 Oct 8;167(2):239–249. [PubMed]
  • Nishikido T, Wada T. Comparative studies of NADP-malic enzyme from C-4 and C-3 plants. Biochem Biophys Res Commun. 1974 Nov 6;61(1):243–249. [PubMed]
  • Persanov VM, Voronova EA, Oparina LA, Karpilov Iu S. Vydelenie i ochistka NADP-zavisimogo "malik"-enzima iz list'ev kukuruzy. Biokhimiia. 1976 May;41(5):921–925. [PubMed]
  • Schnarrenberger C, Herbert M, Krüger I. Intracellular compartmentation of isozymes of sugar phosphate metabolism in green leaves. Isozymes Curr Top Biol Med Res. 1983;8:23–51. [PubMed]
  • Wedding Randolph T. Malic Enzymes of Higher Plants: Characteristics, Regulation, and Physiological Function. Plant Physiol. 1989 Jun;90(2):367–371. [PubMed]
  • Willeford Kenneth O, Wedding Randolph T. pH Effects on the Activity and Regulation of the NAD Malic Enzyme. Plant Physiol. 1987 Aug;84(4):1084–1087. [PubMed]
  • Winter Klaus, Foster Joyce G, Edwards Gerald E, Holtum Joseph A M. Intracellular Localization of Enzymes of Carbon Metabolism in Mesembryanthemum crystallinum Exhibiting C(3) Photosynthetic Characteristics or Performing Crassulacean Acid Metabolism. Plant Physiol. 1982 Feb;69(2):300–307. [PubMed]
  • Wise LS, Rubin CS. Rapid purification and radioimmunoassay of cytosolic malic enzyme. Anal Biochem. 1984 Jul;140(1):256–263. [PubMed]
Articles from Plant Physiology are provided here courtesy of
American Society of Plant Biologists

PubMed articles by these authors