Differential exposure of tryptophan residues in the red and far-red light absorbing forms of phytochrome, as revealed by chemical modification

Miki Nakazawa, Katsushi Manabe

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

The effects of the chemical modification of tryptophan residues in native pea (Pisum sativum L.) phytochrome by 2-hydroxy-5-nitrobenzyl bromide (HNB-Br) were examined. Such treatment had no effect on the spectral properties or on the pattern of tryptic digestion of phytochrome, which indicated that no major conformational change in phytochrome had occurred. Amino acid analysis of the HNB-Br-treated phytochrome indicated that the number of modified Trp residues after the treatment was dependent on the light-absorbing form. The values were three for PR and five for PFR (out of a total of ten) per monomer. The results indicate that two additional Trp residues are exposed on the molecular surface of PFR when the photoconversion of PR to PFR occurs. The amino acid analysis of a 58-kDa tryptic fragment of phytochrome (a mixture of peptides, residues 63-583 and 66-587) showed that one Trp residue in the fragment from PR and two in that from PFR (out of six) were modified by HNB-Br. In the 56-kDa fragment (a mixture of peptides, residues 598-1121 and 603-1124), there were two modified Trp residues in PR and three in PFR (out of four). The Trp residue in a 36-kDa fragment (residues 66-383), which includes the tetrapyrrolic chromophore, was not modified in the either case. These results indicate that new exposed sites that are generated by the photoconversion of PR to PFR are in the region between Trp-456 and Trp-567 and in that between Trp-644 and Trp-787.

Original languageEnglish
Pages (from-to)1097-1105
Number of pages9
JournalPlant and Cell Physiology
Volume34
Issue number7
DOIs
Publication statusPublished - Oct 1993
Externally publishedYes

Keywords

  • 2-Hydroxy-5-nitrobenzyl bromide
  • Chemical modification of tryptophan
  • Limited proteolysis
  • Phytochrome
  • Pisum sativum
  • Tryptophan-chromophore interaction

Field of Science

  • 1.6 Biological sciences
  • 1.4 Chemical sciences

Publication Type

  • 1.1. Scientific article indexed in Web of Science and/or Scopus database

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