Use of fluorescence probes 1-aniline-8-naphthalene sulfonate and pyrene for studying the localisation of proteins in inner membranes from wheat etioplasts

• Published in: Photosynthetica Vol. 33; no. 2
• Main Authors: Denev, I.D, Minkov, I.N. (University of Plovdiv (Bulgaria). Plant Physiology Dept.)
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Nature B.V 01-01-1997
• Subjects: AHILAMIENTO; Aniline; CHLOROPHYLLE; CHLOROPHYLLS; CLOROFILAS; DARKNESS; ETIOLATION; ETIOLEMENT; Etioplasts; FLUORESCENCE; FLUORESCENCIA; Fluorescent indicators; Inner membranes; Irradiation; Lipid bilayers; LIPIDE; LIPIDOS; LIPIDS; Localization; Membrane proteins; Membranes; NADPH-protochlorophyllide oxidoreductase; Naphthalene; OBSCURIDAD; OBSCURITE; OXIDOREDUCTASES; OXIDORREDUCTASAS; OXYDOREDUCTASE; Probes; Proteins; Protochlorophyllide; Pyrene; Quenching; Radiation; Seedlings; Sulfonates; TRIPTOFANO; TRITICUM AESTIVUM; TRYPTOPHAN; TRYPTOPHANE; Wheat
• ISSN: 0300-3604; 1573-9058
• DOI: 10.1023/A:1022180817695

The fluorescence probes 1-aniline-8-naphthalene sulfonate (ANS) and pyrene were applied for characterisation of the light-induced changes in etioplast inner membranes (EPIMs) from 7 d-old dark-grown wheat seedlings (Triticum aestivum L. cv. Pobeda). The major aim was to obtain information about the localisation of membrane proteins in the EPIMs, using probes situated in different regions of the membranes. The quenching of tryptophan fluorescence showed tha the main parts of proteins were accessible to the pyrene buried in the lipid bilayer which suggests that most of the proteins also enter the lipid bilayer. The substantial quenching of the tryptophan fluorescence by the surface-situated ANS demonstrated that a part of the tryptophan residues was probably localised close to the membrane surface. The registered changes after irradiation could be explained by the presence of large aggregates of NADPH-protochlorophyllide oxidoreductase (POR), protochlorophyllide (PChlide) and NADPH in membranes that start to disconnect and redistribute along the prothylakoids.