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Kaiser G, Martinoia E, Schmitt JM, et al. Polypeptide pattern and enzymic character of vacuoles isolated from barley mesophyll protoplasts. Planta. 1986;169(3):345-55doi: 10.1007/BF00392130.
Kaiser, G., Martinoia, E., Schmitt, J. M., Hincha, D. K., & Heber, U. (1986). Polypeptide pattern and enzymic character of vacuoles isolated from barley mesophyll protoplasts. Planta, 169(3), 345-55. https://doi.org/10.1007/BF00392130
Kaiser, G, et al. "Polypeptide pattern and enzymic character of vacuoles isolated from barley mesophyll protoplasts." Planta vol. 169,3 (1986): 345-55. doi: https://doi.org/10.1007/BF00392130
Kaiser G, Martinoia E, Schmitt JM, Hincha DK, Heber U. Polypeptide pattern and enzymic character of vacuoles isolated from barley mesophyll protoplasts. Planta. 1986 Nov;169(3):345-55. doi: 10.1007/BF00392130. PMID: 24232646.
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Planta. 1986 Nov;169(3):345-55. doi: 10.1007/BF00392130.

Polypeptide pattern and enzymic character of vacuoles isolated from barley mesophyll protoplasts.

Planta

G Kaiser, E Martinoia, J M Schmitt, D K Hincha, U Heber

Affiliations

  1. Lehrstuhl Botanik I der Universität Würzburg, Mittlerer Dallenbergweg 64, D-8700, Würzburg, Federal Republic of Germany.

PMID: 24232646 DOI: 10.1007/BF00392130

Abstract

Intact chloroplasts and vacuoles were isolated from mesophyll protoplasts of barley. The chloroplasts occupied about 15% of the cellular volume and contained 75% of the protein, whereas the vacuoles occupied about 80% of the volume and contained less than 4% of total cellular protein. Contamination of the vacuolar fraction by foreign protein is included in these values. Chlorophyll was absent from the vacuolar fraction, but less than 1% of several extra-vacuolar marker proteins were still present. The vacuoles contained hydrolytic enzymes. Several of them (α-mannosidase, α-galactosidase, N-acetylglucosaminidase) were soluble, whereas part of the activity of others semimented with the tonoplasts during centrifugation. Attached proteins could be released from the membranes during freezing in the presence of NaCl. One-dimensional gel electrophoretic separation of soluble vacuolar proteins under non-denaturing conditions yielded more than 10 protein bands. A comparative analysis was performed of thylakoids and vacuoles which were subfractionated into tonoplasts and soluble vacuolar constituents. Sodium dodecyl sulfate gel electrophoresis separated about 15 polypeptides of the soluble fraction which reacted with silver reagent. The tonoplast fraction yielded about 20 bands. A similar number of bands was observed when vacuoles incubated with the (14)C-labelled SH-reagent N-ethylmaleimide were analysed for radioactive polypeptides. Silverstaining of the polypeptides and their SH-content did not correlate. Several polypeptides of the vacuolar fraction had molecular weights very similar to the molecular weights of known chloroplast proteins. However, with the exception of the two subunits of ribulose-1,5-bisphosphate carboxylase, contamination of the vacuolar fraction by chloroplast proteins could be ruled out as a possible cause of the close correspondence. The lipophilic carboxylic-group reagent N,N'-dicyclohexylcarbodiimide ([(14)C]DCCD) reacted with several polypeptides of thylakoids and tonoplasts. However, the labelling patterns were different. The most heavily labelled polypeptide of thylakoids was the 8-kDa polypeptide of the basal part of the coupling factor CF0. Tonoplast polypeptides heavily labelled with [(14)C]DCCD had molecular weights of 24, 28, and 56 kDa. The vacuolar 8-kDa polypeptide remained unlabelled.

References

  1. Biochim Biophys Acta. 1984 Dec 17;768(3-4):209-26 - PubMed
  2. J Cell Biol. 1980 Oct;87(1):72-83 - PubMed
  3. Proc Natl Acad Sci U S A. 1966 Oct;56(4):1095-101 - PubMed
  4. Eur J Biochem. 1974 Jul 1;46(1):83-8 - PubMed
  5. Biochim Biophys Acta. 1978 Oct 11;504(1):67-83 - PubMed
  6. Plant Physiol. 1976 May;57(5):673-80 - PubMed
  7. J Biol Chem. 1982 Jan 25;257(2):1095-101 - PubMed
  8. Anal Biochem. 1978 May;86(1):142-6 - PubMed
  9. Plant Physiol. 1981 Dec;68(6):1354-8 - PubMed
  10. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
  11. Planta. 1984 Nov;161(6):562-8 - PubMed
  12. Plant Physiol. 1979 Jun;63(6):1123-32 - PubMed
  13. Plant Physiol. 1985 Dec;79(4):1090-3 - PubMed
  14. Eur J Biochem. 1979 Sep;99(2):253-9 - PubMed
  15. Plant Physiol. 1986 Apr;80(4):895-901 - PubMed
  16. Methods Enzymol. 1974;32:392-406 - PubMed
  17. Plant Physiol. 1984 Jul;75(3):542-7 - PubMed
  18. J Biol Chem. 1951 Nov;193(1):265-75 - PubMed
  19. Plant Physiol. 1949 Jan;24(1):1-15 - PubMed

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