Measurement of Phytochrome B Thermal Reversion Rates In Vivo.

In vivo spectroscopy Phytochrome Ratiospectrophotometer Thermal reversion

Journal

Methods in molecular biology (Clifton, N.J.)

ISSN: 1940-6029

Titre abrégé: Methods Mol Biol

Pays: United States

ID NLM: 9214969

Informations de publication

Date de publication:
2024

Historique:

medline: 10 4 2024

pubmed: 10 4 2024

entrez: 9 4 2024

Statut: ppublish

Résumé

Thermal reversion of phytochromes is the light-independent but strongly temperature-dependent relaxation of the light-activated Pfr form of phytochromes back into the inactive Pr ground state. The thermal reversion rates of different phytochromes vary considerably. For phytochrome B (phyB), thermal reversion represents a critical parameter affecting phyB activity as it reduces the active phyB Pfr pool, accelerated by increasing temperatures. Phytochromes are dimers existing in three different states: Pfr-Pfr homodimer, Pfr-Pr heterodimer, and Pr-Pr homodimer. Consequently, thermal reversion occurs in two steps, with Pfr-Pfr to Pfr-Pr reversion being much slower than reversion from Pfr-Pr to Pr-Pr. To measure thermal reversion in vivo, the relative proportion of Pfr in relation to the total amount of phytochrome (Ptot) must be determined in living samples. This is accomplished by in vivo spectroscopy utilizing dual wavelength ratiospectrophotometers, optimized for assaying phytochromes in highly scattering plant material. The method is depending on the photoreversibility of phytochromes displaying light-induced absorbance changes in response to actinic irradiation. In this chapter, we describe the experimental design and explain step-by-step the calculations necessary to determine the thermal reversion rates of phyB in vivo, taking into account phytochrome dimerization.

Identifiants

DOI: 10.1007/978-1-0716-3814-9_9 PMID: 38594530

pubmed: 38594530

doi: 10.1007/978-1-0716-3814-9_9

doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

Pagination

85-93

Informations de copyright

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