Nearest neighbor relationships among constituent proteins of oxygen-evolving Photosystem II membranes

binding and function of the extrinsic 33 kDa protein

Isao Enami, Takeshi Miyaoka, Yasuki Mochizuki, Jian-Ren Shen, Kazuhiko Satoh, Sakae Katoh

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Nearest neighbor relationships among constituent proteins of oxygen-evolving Photosystem II (PS II) membrane preparations from spinach were investigated by means of crosslinking with a cleavable bifunctional crosslinker, dithiobis(succinimidylpropionate) (DSP). (1) Diagonal gel electrophoresis revealed crosslinking between two extrinsic proteins of 17 and 23 kDa, between the extrinsic 33 kDa protein and the 47 kDa chlorophyll-carrying protein and between the 26 and 27 kDa apoproteins of light-harvesting chlorophyll a/b protein. In addition, a product which involved a protein of 29 kDa was detected. (2) Amounts of the extrinsic proteins crosslinked were determined by washing DSP-treated membranes with high concentrations of urea and NaCl, or CaCl2. Neither of the two extrinsic proteins of 17 and 23 kDa was crosslinked with intrinsic membrane proteins, whereas 15 to 20% of the 33 kDa protein was immobilized by treatment with 0.1% DSP. Oxygen evolution became resistant to the urea/NaCl-wash proportionally to the amount of the 33 kDa protein crosslinked, indicating that the 33 kDa protein covalently bound to the 47 kDa protein is still fully functional. (3) The crosslinking of the 33 kDa protein accompanied by parallel increases in the amount of Mn remained unextracted, and the rate of oxygen evolution survived after 30-min treatment of PS IImembranes at pH 9.0. Thus the 33 kDa protein has a protective effect on the Mn cluster at the alkaline pH. In contrast, Mn was mostly extracted with a high concentration of Tris, irrespective of the crosslinking of the 33 kDa protein.

Original languageEnglish
Pages (from-to)35-40
Number of pages6
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume973
Issue number1
DOIs
Publication statusPublished - 1989
Externally publishedYes

Fingerprint

Photosystem II Protein Complex
Oxygen
Membranes
Proteins
Crosslinking
Urea
Chlorophyll Binding Proteins
Immobilized Proteins
Apoproteins
Spinacia oleracea
Chlorophyll
Electrophoresis
Washing
Membrane Proteins

Keywords

  • Crosslinking
  • Extrinsic 33 kDa protein
  • Mn cluster
  • Nearest neighbor relationship
  • Oxygen evolution

ASJC Scopus subject areas

  • Biophysics

Cite this

Nearest neighbor relationships among constituent proteins of oxygen-evolving Photosystem II membranes : binding and function of the extrinsic 33 kDa protein. / Enami, Isao; Miyaoka, Takeshi; Mochizuki, Yasuki; Shen, Jian-Ren; Satoh, Kazuhiko; Katoh, Sakae.

In: Biochimica et Biophysica Acta - Bioenergetics, Vol. 973, No. 1, 1989, p. 35-40.

Research output: Contribution to journalArticle

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abstract = "Nearest neighbor relationships among constituent proteins of oxygen-evolving Photosystem II (PS II) membrane preparations from spinach were investigated by means of crosslinking with a cleavable bifunctional crosslinker, dithiobis(succinimidylpropionate) (DSP). (1) Diagonal gel electrophoresis revealed crosslinking between two extrinsic proteins of 17 and 23 kDa, between the extrinsic 33 kDa protein and the 47 kDa chlorophyll-carrying protein and between the 26 and 27 kDa apoproteins of light-harvesting chlorophyll a/b protein. In addition, a product which involved a protein of 29 kDa was detected. (2) Amounts of the extrinsic proteins crosslinked were determined by washing DSP-treated membranes with high concentrations of urea and NaCl, or CaCl2. Neither of the two extrinsic proteins of 17 and 23 kDa was crosslinked with intrinsic membrane proteins, whereas 15 to 20{\%} of the 33 kDa protein was immobilized by treatment with 0.1{\%} DSP. Oxygen evolution became resistant to the urea/NaCl-wash proportionally to the amount of the 33 kDa protein crosslinked, indicating that the 33 kDa protein covalently bound to the 47 kDa protein is still fully functional. (3) The crosslinking of the 33 kDa protein accompanied by parallel increases in the amount of Mn remained unextracted, and the rate of oxygen evolution survived after 30-min treatment of PS IImembranes at pH 9.0. Thus the 33 kDa protein has a protective effect on the Mn cluster at the alkaline pH. In contrast, Mn was mostly extracted with a high concentration of Tris, irrespective of the crosslinking of the 33 kDa protein.",
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