Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1996 Oct 15;319(Pt 2):585–589. doi: 10.1042/bj3190585

Structural changes in photosystem II after treatment with the zero-length bifunctional cross-linker 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide: an electron microscopic study.

R F Collins 1, T D Flint 1, A Holzenburg 1, R C Ford 1
PMCID: PMC1217807  PMID: 8912698

Abstract

Two-dimensional (2D) crystals of photosystem II (PS II) treated with various concentrations of the zero-length crosslinker 1-ethyl-3-(3-dimethylaminopropyl))carbodi-imide (EDC) were analysed by electron microscopy in conjunction with crystallographic image processing. The preparations were characterized by SDS/PAGE and oxygen-evolution measurements, and the effectiveness of cross-linking was monitored by measuring the level of protection afforded against high concentrations of NaCl and CaCl2, which normally remove extrinsic proteins from PS II. We found that low concentrations of EDC (0.25%) increase the order of 2D crystals of PS II. Treatments with EDC concentrations higher than 0.5% did not improve the order of 2D crystals but induced gross structural changes, which were correlated with a decrease in oxygen evolution activity. Structural changes due to cross-linking did not affect packing or symmetry of the 2D crystals, further supporting the conclusion that PS II has a monomeric nature in vivo.

Full Text

The Full Text of this article is available as a PDF (522.4 KB).

Selected References

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

  1. Amos L. A., Henderson R., Unwin P. N. Three-dimensional structure determination by electron microscopy of two-dimensional crystals. Prog Biophys Mol Biol. 1982;39(3):183–231. doi: 10.1016/0079-6107(83)90017-2. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Bassi R., Ghiretti Magaldi A., Tognon G., Giacometti G. M., Miller K. R. Two-dimensional crystals of the photosystem II reaction center complex from higher plants. Eur J Cell Biol. 1989 Oct;50(1):84–93. [PubMed] [Google Scholar]
  4. Boekema E. J., Boonstra A. F., Dekker J. P., Rögner M. Electron microscopic structural analysis of Photosystem I, Photosystem II, and the cytochrome b6/f complex from green plants and cyanobacteria. J Bioenerg Biomembr. 1994 Feb;26(1):17–29. doi: 10.1007/BF00763217. [DOI] [PubMed] [Google Scholar]
  5. Bricker T. M., Frankel L. K. Use of a monoclonal antibody in structural investigations of the 49-kDa polypeptide of photosystem II. Arch Biochem Biophys. 1987 Jul;256(1):295–301. doi: 10.1016/0003-9861(87)90449-8. [DOI] [PubMed] [Google Scholar]
  6. Green B. R., Pichersky E., Kloppstech K. Chlorophyll a/b-binding proteins: an extended family. Trends Biochem Sci. 1991 May;16(5):181–186. doi: 10.1016/0968-0004(91)90072-4. [DOI] [PubMed] [Google Scholar]
  7. Jansson S. The light-harvesting chlorophyll a/b-binding proteins. Biochim Biophys Acta. 1994 Feb 8;1184(1):1–19. doi: 10.1016/0005-2728(94)90148-1. [DOI] [PubMed] [Google Scholar]
  8. Kühlbrandt W. Two-dimensional crystallization of membrane proteins. Q Rev Biophys. 1992 Feb;25(1):1–49. doi: 10.1017/s0033583500004716. [DOI] [PubMed] [Google Scholar]
  9. Lyon M. K., Marr K. M., Furcinitti P. S. Formation and characterization of two-dimensional crystals of photosystem II. J Struct Biol. 1993 Mar-Apr;110(2):133–140. doi: 10.1006/jsbi.1993.1014. [DOI] [PubMed] [Google Scholar]
  10. Masojidek J., Droppa M., Horváth G. Analysis of the polypeptide composition of grana and stroma thylakoids by two-dimensional gel electrophoresis. Distribution of photosystem II between grana and stroma lamellae. Eur J Biochem. 1987 Dec 1;169(2):283–288. doi: 10.1111/j.1432-1033.1987.tb13609.x. [DOI] [PubMed] [Google Scholar]
  11. Miller K. R., Cushman R. A. A chloroplast membrane lacking photosystem II. Thylakoid stacking in the absence of the photosystem II particle. Biochim Biophys Acta. 1979 Jun 5;546(3):481–497. doi: 10.1016/0005-2728(79)90083-5. [DOI] [PubMed] [Google Scholar]
  12. Nicholson W. V., Ford R. C., Holzenburg A. A current assessment of photosystem II structure. Biosci Rep. 1996 Apr;16(2):159–187. doi: 10.1007/BF01206204. [DOI] [PubMed] [Google Scholar]
  13. Preston C., Seibert M. The carboxyl modifier 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC) inhibits half of the high-affinity Mn-binding site in photosystem II membrane fragments. Biochemistry. 1991 Oct 8;30(40):9615–9624. doi: 10.1021/bi00104a008. [DOI] [PubMed] [Google Scholar]
  14. Santini C., Tidu V., Tognon G., Ghiretti Magaldi A., Bassi R. Three-dimensional structure of the higher-plant photosystem II reaction centre and evidence for its dimeric organization in vivo. Eur J Biochem. 1994 Apr 1;221(1):307–315. doi: 10.1111/j.1432-1033.1994.tb18742.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES