Differences between the surface properties of the PR and PFR forms of native pea phytochrome, and their application to a simplified procedure for purification of the phytochrome

Miki Nakazawa, Yuhji Yoshida, Katsushi Manabe

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

The ability of phytochrome from etiolated pea shoots (Pisum sativum L. cv. Alaska) to bind to various chromatographic adsorbents and its mobility during non-denaturing electrophoresis were examined with phytochrome in either the red light-absorbing form (PR) or the far-red light-absorbing form (PFR). Preferential binding of PFR to modified hydrophilic polyvinyl resins, such as butyl Toyopearl, phenyl Toyopearl, Blue Toyopearl (Cibacron Blue F3G-A conjugated) and Red Toyopearl (Procion Red HE-3B conjugated), was observed. A simplified method for purification of native phytochrome was developed based on the properties of PR and PFR. PFR bound preferentially to the hydrophobic adsorbents, to indicate that the surface of PFR is more hydrophobic than that of PR. A difference in net surface charges between PR and PFR was detected by an analysis based on the different mobilities of the two forms during non-denaturing polyacrylamide gel electrophoresis in gels prepared with various concentrations of polyacrylamide. The apparent molecular weights of PR and PFR, estimated from the analysis, were 378 and 419 kilodaltons, respectively. The difference suggests that a significant change in molecular shape occurs during the photoconversion. The differences in surface properties of PR and PFR are discussed.

Original languageEnglish
Pages (from-to)1187-1194
Number of pages8
JournalPlant and Cell Physiology
Volume32
Issue number8
DOIs
Publication statusPublished - Dec 1991
Externally publishedYes

Keywords

  • Ferguson plot analysis
  • Hydrophobic chromatography
  • Phytochrome
  • Pisum sativum
  • Procion Red HE-3B affinity
  • Purification

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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