Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Jun;78(6):3595–3599. doi: 10.1073/pnas.78.6.3595

Separation and characterization of inner and outer envelope membranes of pea chloroplasts

Kenneth Cline 1, Jaen Andrews 1, Brent Mersey 1, Eldon H Newcomb 1, Kenneth Keegstra 1
PMCID: PMC319617  PMID: 16593034

Abstract

A procedure for purifying the chloroplast envelope subfractionates it into two membrane fractions of comparable quantities. This procedure differs from previous ones in that the chloroplasts are ruptured by freezing and thawing in hypertonic medium rather than by osmotic shock. The two membrane fractions have qualitatively similar polar lipid compositions but differ in their content of individual lipids, specifically monogalactosyldiacylglycerol and phosphatidylcholine. The two fractions also differ in their constituent polypeptides and in their appearance when examined by electron microscopy. The light (density = 1.08 g/ml) and heavy (density = 1.13 g/ml) membrane fractions have been tentatively identified as the outer and inner envelope membranes, respectively.

Keywords: freeze-thaw hypertonic lysis, flotation centrifugation, galactolipids, galactosyl transferase, protease-treated chloroplasts

Full text

PDF
3595

Images in this article

3597Image
on p.3597
3598Image
on p.3598
3598Image
on p.3598

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chua N. H., Schmidt G. W. Transport of proteins into mitochondria and chloroplasts. J Cell Biol. 1979 Jun;81(3):461–483. doi: 10.1083/jcb.81.3.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Decker G. L., Greenawalt J. W. Ultrastructural and biochemical studies of mitoplasts and outer membranes derived from French-pressed mitochondria. Advances in mitochondrial subfractionation. J Ultrastruct Res. 1977 Apr;59(1):44–56. doi: 10.1016/s0022-5320(77)80027-0. [DOI] [PubMed] [Google Scholar]
  4. Douce R., Mannella C. A., Bonner W. D., Jr The external NADH dehydrogenases of intact plant mitochondria. Biochim Biophys Acta. 1973 Jan 18;292(1):105–116. doi: 10.1016/0005-2728(73)90255-7. [DOI] [PubMed] [Google Scholar]
  5. GERLACH E., DEUTICKE B. EINE EINFACHE METHODE ZUR MIKROBESTIMMUNG VON PHOSPHAT IN DER PAPIERCHROMATOGRAPHIE. Biochem Z. 1963 Jul 26;337:477–479. [PubMed] [Google Scholar]
  6. Heldt H. W., Sauer F. The inner membrane of the chloroplast envelope as the site of specific metabolite transport. Biochim Biophys Acta. 1971 Apr 6;234(1):83–91. doi: 10.1016/0005-2728(71)90133-2. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  9. Poincelot R. P., Day P. R. An improved method for the isolation of spinach chloroplast envelope membranes. Plant Physiol. 1974 Nov;54(5):780–783. doi: 10.1104/pp.54.5.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Spurr A. R. A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res. 1969 Jan;26(1):31–43. doi: 10.1016/s0022-5320(69)90033-1. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES