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Lily cofactor-independent phosphoglycerate mutase: purification, partial sequencing, and immunolocalization

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Abstract

A cofactor-independent phosphoglycerate mutase (PGAM-i) was isolated to homogeneity from monocotyledonous Lilium longiflorum Thunb. Two-dimensional (2D) polyacrylamide gel electrophoresis resolved three PGAM-i forms. This enzyme was originally identified by cross-reactivity to antibodies affinity-purified from 2D gels using human vitronectin (VN). Antibody produced against a denatured protein spot from a 2D gel did not recognize VN protein, but partial protein and DNA sequencing showed similarity of the former protein to maize PGAM-i. Immunoblots from roots, styles, leaves, and anthers showed the presence of PGAM-i in all tissues examined; it was isolated predominantly from the soluble cell fraction, with some present in the insoluble cell fraction. Immunoelectron microscopy demonstrated its localization in the cytoplasm and plastids in root cells near the apical meristem. In addition, immunogold labeling detected signals from the nucleus. The immunohistochemical localization of the enzyme in the nucleus, as well as in the cytosol and plastids, indicates that lily PGAM-i might have multiple functions in the cell.

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Abbreviations

ELISA:

enzyme-linked immunosorbent assay

PGAM:

cofactor-dependent phosphoglycerate mutase

PGAM-i:

cofactor independent phosphoglycerate mutase

TEM:

transmission electron microscopy

2D:

two dimensional

VN:

vitronectin

References

  • Balsamo RA, Wang J-L, Eckard KJ, Wang C-S, Lord EM (1995) Immunogold localization of a developmentally regulated, tapetal-specific, 15-kDa lily protein. Protoplasma 189: 17–25

    Google Scholar 

  • Botha FC, Dennis DT (1986) Isozymes of phosphoglyceromutase from the developing endosperm of Ricinus communis: Isolation and kinetic properties. Arch Biochem Biophys 245: 96–103

    Google Scholar 

  • Botha FC, Dennis DT (1987) Phosphoglyceromutase activity and concentration in the endosperm of developing and germinating Ricinus communis seeds. Can J Bot 65: 1908–1912

    Google Scholar 

  • Britton HG, Carreras J, Grisolia S (1971) Mechanism of action of 2,3-diphosphoglycerate-independent phosphoglycerate mutase. Biochemistry 10: 4522–4533

    Google Scholar 

  • Carreras J, Mezquita J, Bosch J, Bartrons GP (1982) Phylogeny and ontogeny of the phosphoglycerate mutases — IV. Distribution of glycerate-2,3-P2 dependent and independent phosphoglycerate mutases in algae, fungi, plants and animais. Comp Biochem Physiol 71: 591–597

    Google Scholar 

  • Chang YC, Walling LL (1992) Spatial and temporal expression of Cab mRNAs in cotyledons of the developing soybean seedling. Planta 186: 262–272

    Google Scholar 

  • Copeland L, Turner JF (1987) The regulation of glycolysis and the pentose phosphate pathway. In: Davies DD (ed) The biochemistry of plants. A comprehensive treatise, vol 11. Biochemistry of metabolism. Academic Press, San Diego, pp 107–128

    Google Scholar 

  • Dennis DT, Miernyk JA (1982) Compartmentation of nonphotosynthetic carbohydrate metabolism. Annu Rev Plant Physiol 33: 27–50

    Google Scholar 

  • Egea G, Urena JM, Grana X, Marsal J, Carreras J, Climent F (1992) Nuclear location of phosphoglycerate mutase BB isozyme in rat tissues. Histochemistry 97: 269–275

    Google Scholar 

  • Engvall E, Perlmann P (1972) Enzyme-linked immunosorbent assay, ELISA. III. Quantitation of specific antibodies by enzymelabeled anti-immunoglobulin in antigen-coated tubes. J Immunol 109: 129–135

    Google Scholar 

  • Faye L, Greenwood JS, Herman EM, Sturm A, Crispeels MJ (1988) Transport and posttranslational processing of the vacuolar enzyme α-mannosidase in jack-bean cotyledons. Planta 174: 271–282

    Google Scholar 

  • Fothergill-Gilmore LA, Watson HC (1989) The phosphoglycerate mutases. Adv Enzymol 62: 227–313

    Google Scholar 

  • Grana X, de Lecea L, El-Maghrabi MR, Urena JM, Caellas C, Carreras J, Puigdomenech P, Pilkis SJ, Climent F (1992) Cloning and sequencing of a cDNA encoding 2,3-bisphosphoglycerate independent phosphoglycerate mutase from maize. J Biol Chem 267: 12797–12803

    Google Scholar 

  • Grana X, Broceno C, Garriga J, Ossa de la P, Climent F (1993) Phosphoglycerate mutase activity and mRNA levels during germination of maize embryos. Plant Sci 89: 147–151

    Google Scholar 

  • Hakes DJ, Berezney R (1991) Molecular cloning of matrix F/G-A DNA binding protein of the nuclear matrix that contain putative zinc finger motifs. Proc Natl Acad Sci USA 88: 6186–6190

    Google Scholar 

  • Henikoff S (1987) Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol 155: 156–165

    Google Scholar 

  • Huang Y, Blakeley SD, McAleese SM, Fothergill-Gilmore LA, Dennis DT (1993) Higher-plant cofactor-independent phosphoglyceromutase: Purification molecular characterization and expression. Plant Mol Biol 23: 1039–1053

    Google Scholar 

  • Jindal HK, Vishwanatha JK (1990) Functional identity of a primer recognition protein as phosphoglycerate kinase. J Biol Chem 265: 6540–6543

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    PubMed  Google Scholar 

  • Manfioletti G, Schneider C (1988) A new and fast method for preparing high quality lambda DNA suitable for sequencing. Nucleic Acids Res 16: 2873–2884

    Google Scholar 

  • Murphy DV, Leech RM (1977) Lipid biosynthesis from (14C)bicarbonate, (2·4C)pyruvate and (I-14C)acetate during photosynthesis by isolated chloroplasts. FEBS Lett 77: 164–168

    Google Scholar 

  • O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021

    Google Scholar 

  • Ossa de la P, Grana X, Ruiz-Lozano P, Climent F (1994) Isolation and characterization of cofactor-independent phosphoglycerate mutase gene from maize. Biochem Biophys Res Commun 203: 1204–1209

    Google Scholar 

  • Piatigorsky J, Wistow GJ (1989) Enzyme/crystallins: gene sharing as an evolutionary strategy. Cell 57: 197–199

    Google Scholar 

  • Pautot V, Holzer FM, Walling LL (1991) Expression of proteinase inhibitor genes during Pseudomonas syrinyae pv. tomato infection. Mol Plant Microb Interact 4: 284–292

    Google Scholar 

  • Polson A (1990) Isolation of IgY from the yolks of eggs by a chloroform polyethylene glycol procedure. Immun Invest 19: 253–258

    Google Scholar 

  • Sanders LC, Wang C-S, Walling LL, Lord EM (1991) A homolog of the substrate adhesion molecule vitronectin occurs in four species of flowering plants. Plant Cell 3: 629–635

    Google Scholar 

  • Simcox PD, Reid EE, Canvin DT, Dennis DT (1977) Enzymes of the glycolytic and pentose phosphate pathways in proplastids from the developing endosperm of Ricinus communis L. Plant Physiol 59: 1128–1132

    Google Scholar 

  • Stitt M, apRees T (1979) Capacities of pea chloroplasts to catalyse the oxidative pentose phosphate pathway and glycolysis. Phytochemistry 18: 1905–1911

    Google Scholar 

  • Urena JM, Egea G, Grana X, Castella J, Marsal J, Carreras J, Climent F (1990) Location of phosphoglycerate mutase in rat skeletal muscle. An immunocytochemical and biochemical study. Eur J Cell Biol 51: 151–156

    Google Scholar 

  • Wang C-S, Walling LL, Eckard KJ, Lord EM (1992) Patterns of protein accumulation in developing anthers of Lilium longiflorum correlate with histological events. Am J Bot 79: 118–127

    Google Scholar 

  • White MF, Fothergill-Gilmore LA (1992) Development of a mutagenesis, expression and purification system for yeast phosphoglycerate mutase. Investigation of the role of active-site His181. Eur J Biochem 207: 709–714

    Google Scholar 

  • Yamada M, Nakamura Y (1975) Fatty acid synthesis by spinach chloroplasts. II. The path from PGA to fatty acids. Plant Cell Physiol 16: 151–162

    Google Scholar 

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Author notes

  1. J.-L. Wang

    Present address: Department of Surgery, UCLA Medical Center, 10833 Le Conte Ave., B2-394 CHS, 90024-1741, Los Angeles, CA, USA

Authors and Affiliations

  1. Department of Botany and Plant Sciences, University of California, 92521-0124, Riverside, CA, USA

    J.-L. Wang, L. L. Walling, G. Y. Jauh, Y.-Q. Gu & E. M. Lord

Authors
  1. J.-L. Wang
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  2. L. L. Walling
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  3. G. Y. Jauh
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  4. Y.-Q. Gu
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  5. E. M. Lord
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Correspondence to E. M. Lord.

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Wang, JL., Walling, L.L., Jauh, G.Y. et al. Lily cofactor-independent phosphoglycerate mutase: purification, partial sequencing, and immunolocalization. Planta 200, 343–352 (1996). https://doi.org/10.1007/BF00200302

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  • DOI: https://doi.org/10.1007/BF00200302

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