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Planta

, Volume 178, Issue 2, pp 147–156 | Cite as

Phytochrome structure: Peptide fragments from the amino-terminal domain involved in protein-chromophore interactions

  • Alan M. Jones
  • Peter H. Quail
Article

Abstract

We have undertaken a study of the structure of the amino-terminal domain of the phytochrome polypeptide purified from Avena sativa L. Amino-acid sequencing was used to indentify arginine 52 as the precise location of a conformation-specific cleavage of phytochrome by subtilisin. The location of the epitopes for a class of monoclonal antibodies designated type 2 has been shown to be located between approx. 10 and 20 kilodaltons (kDa) from the amino terminus. These two new spatial markers, in addition to the chromophore and another epitope recognized by type 1 monoclonal antibodies and located within 6 kDa from the amino terminus, have been used to map the locations of several new protease-accessible sites along the polypeptide. After extensive digestion of phytochrome with subtilisin, a stable spectrally-active group of peptides remains. Within this group is a 16-kDa chromopeptide which, either alone or as part of an assemblage of peptides, elutes from a size-exclusion column under nondenaturing conditions at a volume consistent with a molecular mass of 35–40 kDa. This group of peptides has an absorbance spectrum similar to the red-absorbing form of phytochrome (Pr) and is red/far-red photoreversible between this and a photobleached form. These data indicate that this group of peptides still retains the principal structural requisites for Pr-chromophore-protein interactions and for photoreversibility, but not for Pfr (far-red-absorbing phytochrome)-chromophore-protein interactions. It is uncertain if these structural requisites reside exclusively on the 16-kDa chromopeptide or result from an assemblage of these peptides. However, we have excluded any role for an adjacent 14-kDa fragment (approximately residues 50 to 200) in the observed spectral properties since it can be selectively removed without any effect on the photoreversibility.

Key words

Avena (phytochrome) Phytochrome (structure, photoreversibility) 

Abbreviations

Da

dalton

Mr

relative molecular mass

Pr, Pfr

red and far-red-absorbing forms of phytochrome, respectively

SDS-PAGE

sodium dodecyl sulfate polyacrylamide gel electrophoresis

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Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Alan M. Jones
    • 1
  • Peter H. Quail
    • 2
  1. 1.Department of BiologyUniversity of North CarolinaChapel HillUSA
  2. 2.Department of BotanyUniversity of WisconsinMadisonUSA

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