Display options
Cite
Arvidsson PO, Bratt CE, Carlsson M, et al. Purification and identification of the violaxanthin deepoxidase as a 43 kDa protein. Photosynth Res. 1996;49(2):119-29doi: 10.1007/BF00117662.
Arvidsson, P. O., Bratt, C. E., Carlsson, M., & Akerlund, H. E. (1996). Purification and identification of the violaxanthin deepoxidase as a 43 kDa protein. Photosynthesis research, 49(2), 119-29. https://doi.org/10.1007/BF00117662
Arvidsson, P O, et al. "Purification and identification of the violaxanthin deepoxidase as a 43 kDa protein." Photosynthesis research vol. 49,2 (1996): 119-29. doi: https://doi.org/10.1007/BF00117662
Arvidsson PO, Bratt CE, Carlsson M, Akerlund HE. Purification and identification of the violaxanthin deepoxidase as a 43 kDa protein. Photosynth Res. 1996 Aug;49(2):119-29. doi: 10.1007/BF00117662. PMID: 24271609.
Copy Download .nbib Format: NLM
Share it on

Photosynth Res. 1996 Aug;49(2):119-29. doi: 10.1007/BF00117662.

Purification and identification of the violaxanthin deepoxidase as a 43 kDa protein.

Photosynthesis research

P O Arvidsson, C E Bratt, M Carlsson, H E Akerlund

Affiliations

  1. Department of Plant Biochemistry, Lund University, P.O.B. 117, S-221 00, Lund, Sweden.

PMID: 24271609 DOI: 10.1007/BF00117662

Abstract

Violaxanthin deepoxidase (VDE) has been purified from spinach (Spinacia oleracea) leaves. The purification included differential sonication of thylakoid membranes, differential (NH4)2SO4 fractionation, gel filtration chromatography and finally either hydrophobic interaction chromatography or anion exchange chromatography. A total purification of more than 5000-fold compared to the original thylakoids enabled the identification of a 43 kDa protein as the VDE, in contrast to earlier reported molecular weight of 54-60 kDa. A detailed comparison was made for the VDE activity and polypeptide pattern for the different fractions throughout the purification and the best correlation was always found for the 43 kDa protein. The highest specific activity obtained was 256 μmol g(-1) s(-1) protein, which is at least 10-fold higher than reported earlier. We estimate that there is 1 VDE molecule per 20-100 electron transport chains. The 43 kDa protein was N-terminally sequenced, after protection of cysteine residues with β-mercaptoethanol and iodoacetamid, and a unique sequence of 20 amino acids was obtained. The amino acid composition of the protein revealed a high abundance of charged and polar amino acids and remarkably, 11 cysteine residues. Two other proteins (39.5 kDa and 40 kDa) copurifying with VDE were also N-terminally sequenced. The N-terminal part of the 39.5 kDa protein showed complete sequence identity both with the N-terminal part of cyt b 6 and an internal sequence of polyphenol oxidase.

References

  1. Proc Natl Acad Sci U S A. 1972 Apr;69(4):930-2 - PubMed
  2. Arch Biochem Biophys. 1978 Oct;190(2):514-22 - PubMed
  3. Photosynth Res. 1993 Sep;37(3):217-25 - PubMed
  4. Planta. 1970 Dec;93(4):314-22 - PubMed
  5. Photosynth Res. 1995 Aug;45(2):169-75 - PubMed
  6. Biochim Biophys Acta. 1972 Jun 23;267(3):538-43 - PubMed
  7. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
  8. Photosynth Res. 1990 Mar;23(3):331-43 - PubMed
  9. Eur J Biochem. 1986 Aug 1;158(3):477-82 - PubMed

Publication Types