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. 1994 Jan;104(1):49–57. doi: 10.1104/pp.104.1.49

Purification of the Major Soybean Leaf Acid Phosphatase That Is Increased by Seed-Pod Removal.

P E Staswick 1, C Papa 1, J F Huang 1, Y Rhee 1
PMCID: PMC159161  PMID: 12232060

Abstract

Fruit removal for 5 weeks after flowering increased acid phosphatase activity 10-fold in soybean (Glycine max L. Merr. Var Hobbit) leaves compared with normal seed-pod-bearing plants. The major acid phosphatase activity in leaves was purified over 2700-fold, yielding a single polypeptide of 51 kD with a specific activity of 1353 units/mg protein using p-nitrophenylphosphate as the substrate. Isoelectric focusing demonstrated that the purified protein co-migrated with a majority of the activity that increased in leaves following seed-pod removal. Immunoblot analysis demonstrated that at least part of the increased activity was due to an increased abundance of the phosphatase protein. In situ enzyme activity staining localized most of the total phosphatase activity to vascular tissues, the leaf paraveinal mesophyll cell layer, and the lower epidermis. This distribution and the response to seed-pod removal paralleled previous results for soybean vegetative storage protein (VSP) [alpha] and [beta]. However, in a native polyacrylamide gel the VSP detected by immunological staining of electrophoretically transferred protein did not migrate with the majority of the phosphatase activity. Fractionation of crude leaf protein on concanavalin A-Sepharose yielded a fraction containing 97% of the total VSP but only 0.1% of the total acid phosphatase activity.

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Selected References

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  1. Aarts J. M., Hontelez J. G., Fischer P., Verkerk R., van Kammen A., Zabel P. Acid phosphatase-1(1), a tightly linked molecular marker for root-knot nematode resistance in tomato: from protein to gene, using PCR and degenerate primers containing deoxyinosine. Plant Mol Biol. 1991 Apr;16(4):647–661. doi: 10.1007/BF00023429. [DOI] [PubMed] [Google Scholar]
  2. Andrews D. L., Beames B., Summers M. D., Park W. D. Characterization of the lipid acyl hydrolase activity of the major potato (Solanum tuberosum) tuber protein, patatin, by cloning and abundant expression in a baculovirus vector. Biochem J. 1988 May 15;252(1):199–206. doi: 10.1042/bj2520199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DeWald D. B., Mason H. S., Mullet J. E. The soybean vegetative storage proteins VSP alpha and VSP beta are acid phosphatases active on polyphosphates. J Biol Chem. 1992 Aug 5;267(22):15958–15964. [PubMed] [Google Scholar]
  4. Duff S. M., Lefebvre D. D., Plaxton W. C. Purification and Characterization of a Phosphoenolpyruvate Phosphatase from Brassica nigra Suspension Cells. Plant Physiol. 1989 Jun;90(2):734–741. doi: 10.1104/pp.90.2.734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Duff S. M., Plaxton W. C., Lefebvre D. D. Phosphate-starvation response in plant cells: de novo synthesis and degradation of acid phosphatases. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9538–9542. doi: 10.1073/pnas.88.21.9538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Huang J. F., Bantroch D. J., Greenwood J. S., Staswick P. E. Methyl jasmonate treatment eliminates cell-specific expression of vegetative storage protein genes in soybean leaves. Plant Physiol. 1991 Dec;97(4):1512–1520. doi: 10.1104/pp.97.4.1512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Laine A. C., Faye L. Significant immunological cross-reactivity of plant glycoproteins. Electrophoresis. 1988 Dec;9(12):841–844. doi: 10.1002/elps.1150091210. [DOI] [PubMed] [Google Scholar]
  8. Nishimura M., Beevers H. Hydrolases in vacuoles from castor bean endosperm. Plant Physiol. 1978 Jul;62(1):44–48. doi: 10.1104/pp.62.1.44. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Staswick P. E. Developmental regulation and the influence of plant sinks on vegetative storage protein gene expression in soybean leaves. Plant Physiol. 1989 Jan;89(1):309–315. doi: 10.1104/pp.89.1.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Staswick P. E. Soybean vegetative storage protein structure and gene expression. Plant Physiol. 1988 May;87(1):250–254. doi: 10.1104/pp.87.1.250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ullah A. H., Gibson D. M. Purification and characterization of acid phosphatase from cotyledons of germinating soybean seeds. Arch Biochem Biophys. 1988 Feb 1;260(2):514–520. doi: 10.1016/0003-9861(88)90476-6. [DOI] [PubMed] [Google Scholar]
  12. Williamson V. M., Colwell G. Acid phosphatase-1 from nematode resistant tomato : isolation and characterization of its gene. Plant Physiol. 1991 Sep;97(1):139–146. doi: 10.1104/pp.97.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wittenbach V. A. Effect of pod removal on leaf senescence in soybeans. Plant Physiol. 1982 Nov;70(5):1544–1548. doi: 10.1104/pp.70.5.1544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wittenbach V. A. Purification and characterization of a soybean leaf storage glycoprotein. Plant Physiol. 1983 Sep;73(1):125–129. doi: 10.1104/pp.73.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]

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