Abstract
Two isoforms of phosphoglucomutase (PGM, EC 2.7.5.1) designated as PGM-I and PGM-II were purified from developing seeds of Brassica campestris L to their electrophoretic homogeneity. Both the forms had molecular mass of 59 kD each and were monomeric. PGM-I exhibited maximum activity at pH 7.5, while PGM-II evinced pH optima at 8.25. Both the forms exhibited hyperbolic response towards increasing concentrations of the substrate with Km values of 0.10 mM for PGM-I and 0.12 mM for PGM-II and had absolute requirement for glucose-1,6-P2. Fructose-1,6-P2 and 2,3-diphosphoglyceric acid inhibited the two forms non-competitively, whereas, ribulose-1,5-P2 inhibited only PGM-II, with Ki value of 0.8 mM. ATP inhibited the enzyme uncompetitively with Ki values for 0.26 mM (PGM-I) and 0.12 mM (PGM-11). Use of group specific protease inhibitors indicated Ser, His and Cys to play significant role in catalysis. On the basis of their differential behaviour and kinetic properties, PGM-I and PGM-II may be the forms present in cytosol and leucoplasts, respectively.
References
Manjunath S, Lee CHK, Van Winkle P & Bailey-Serres J, Plant Physiol, 117 (1998) 997.
Hattenbach A & Heineke D, Planta, 207 (1999) 527.
Singh R, Indian J Biochem Biophys, 29 (1992) 1.
Muhlbach H & Schnarrenberger C, Planta, 141 (1978) 65.
Takamiya S & Fukui T, Plant Cell Physiol, 19 (1978) 759.
Sangwan RS & Singh R, Plant Physiol Biochem, 25 (1987) 745.
Esposito S, Bowsher CG, Emes MJ & Tetlow IJ, J Plant Physiol, 154 (1999) 24.
Tetlow IJ, Blissett KJ & Emes MJ, Planta, 204 (1998) 100.
Archana, Sharma N, Malhotra SP & Singh R, J Plant Biol, 26 (1999) 161.
Periappuram C, Steinhauer L, Barton DL, Taylor DC, Chatson B & Zou J, Plant Physio/, 122 (2000) 1193.
MacDonald FD & apRees T, Biochem Biophys Acfa, 755 (1983) 81.
Lowry NJ, Rosebrough NJ, Farr AL & Randall RJ, J Biol Chem, 193 (1951) 265.
Davis BJ, Ann NY Acad Sci, 121 (1964) 404.
Laemmli UK, Nature, 277 (1970) 680.
Pressey R, J Food Sci, 32 (1967) 381.
Small DM & Matheson NK, Phytochemistry: 18 (1979) 1147.
Salvucci ME, Drake RR, Broadbent KP, Haley BE, Hanson KR & McHale NA, Plant Physiol, 93 (1990) 105.
Gardemann A, Schimkat D & Heldt HW, Planta, 168 (1986) 536.
Miles EW, Methods Enzymol, 47 (1977) 431.
Melchior WB & Fahrney D, Biochemistry, 9 (1970) 251.
Dominici P, Tancini B & Voltattorn CB, J Biol Chem, 260 (1985) 10583.
Deluca V & Dennis DT, Plant Physiol, 61 (1978) 1037.
Zimmermann G, Kelly GJ & Latzko E, JBiol Chem, 253 (1978) 5952.
Singal HR, Sheoran IS & Singh R, Plant Physiol, 83 (1987) 1043.
Casey R, Bomoney C, Forster C, Hedley C, Hitchin E & Wang T, J Plant P hysiol, 152 (1998) 636.
Harrison CJ, Hedley CL & Wang TL, Plant J, 13 (1998) 753.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yadav, S.K., Sharma, N.S. & Singh, R. Purification and Characterization of Phosphoglucomutase from Heterotrophic Tissues of Brassica campestris L. J. Plant Biochem. Biotechnol. 11, 133–137 (2002). https://doi.org/10.1007/BF03263151
Received:
Revised:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF03263151