Purification and partial characterization of glutamine synthetase from root nodules of faba vean

Authors

  • J.M. CABA
  • L. GUZMÁN
  • L. ORTEGA
  • C. LLUCH
  • M. LARA
  • F. LIGERO

Keywords:

Vicia faba, Enzyme isoforms, Glutamine synthetase, Nodules. Root

Abstract

Glutamine synthetase (GS, EC 6.1.3 .2) from the plant fraction of Vicia faba L.

nodules was purified to apparent homogeneity using Sepharose-anthranilic acid affinity

chromatography. The enzyme is composed of three 40 kD polypeptides and has a native

molecular weight of 310-330 kD, determined by gel filtration chromatography and

native gradient polyacrylamide gel electrophoresis, respectively, and showed crossreactivity

with antibodies obtained against Phaseolus vulgaris nodule GS. Two isoenzymes,

GSn-1 and Gsn-2, were separated in nodules by anion-exchange chromatography on QSepharose,

while just one form could be identified in roots, all the isoenzymes being

identical in subunit composition and native molecular weight. The transferase to

semibiosynthetic activity ratio of all three GS forms was found to be unusually low.

Glutamine synthetase abundance in nodules was quantified by densitometry, representing

about 5% of total soluble protein extracted. Possible significance of these and other

characteristics of V. faba GS is discussed.

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References

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PACKOVSKY, R. S., FULLER, G.: "Nitrogen assimilation and partitioning in two nitrogen-fixing cultivars of Phaseolus vulgaris L". Plan! Soil ( 1991), 132:139-148.

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CORUZZI, G. M., EDWARDS, J. W., TINGEY, S. V., TSAI, F.-Y., WALKER, E. L. "Glutamine synthetase: Molecular evolution of an eclectic multigene family". In: Molecular basis 01 plant development (1988), 223-232. R. Goldberg (ed.). Alan R. Liss New York.

FORDE, B. J., CULLIMORE, J. V.: "The molecular biology of glutamine synthetase Ijhigher plants". Oxl Surv Plant Mol Biol (1989), 6:257-296.

LARA, M., PORTA, H., PADILLA, J., FOLCH, J. , SÁNCHEZ, F.: "Heterogeneity o glutamine synthetase polypeptides in Phaseolus vulgaris L". Plant Physiol. (1984) 76:1019-1023 .

HERVÁS, A., CABA, J. M., LIGERO, F., LLUCH, C.: "Effect of combined nitroge: supply and nodulation on nitrate reductase activity and growth of pea plants". J Plan Nutr (1991), 14:1319-1330.

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Published

1997-12-20

How to Cite

1.
CABA J, GUZMÁN L, ORTEGA L, LLUCH C, LARA M, LIGERO F. Purification and partial characterization of glutamine synthetase from root nodules of faba vean. Ars Pharm [Internet]. 1997 Dec. 20 [cited 2024 May 19];38(4):345-56. Available from: https://revistaseug.ugr.es/index.php/ars/article/view/21272

Issue

Vol. 38 No. 4 (1997)

Section

Original Articles

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