Molecular rearrangement in POR macrodomains as a reason for the blue shift of chlorophyllide fluorescence observed after phototransformation.
The activation energy and activation volume of the spectral blue shift subsequent to protochlorophyllide phototransformation (called Shibata shift in intact leaves) were studied in prolamellar body (PLB) and prothylakoid-(PT)-enriched membrane fractions prepared from dark-grown wheat (Triticum aestivum, L.) leaves. The measurements were done at 20, 30 and 40 degrees C and at various pressure values. The activation energy values were 181+/-8 kJ mol(-1) and 188+/-6 kJ mol(-1) for the PLBs and the PTs, respectively. The pressure stabilized the structure of the NADPH:protochlorophyllide oxidoreductase (POR) macrodomains; it prevented or slowed down the blue shift. There were no significant differences between the activation volumes of PLBs and PTs at 30 or 40 degrees C giving values around 100-125 ml mol(-1) which correspond to changes in the tertiary structure of proteins but also resemble the volume changes occurring during the disaggregation of protein dimers or oligomers, or during dissociation of peripheral membrane proteins from membranes. The small differences in the activation parameters of PLBs and PTs indicate that molecular rearrangements inside the POR macrodomains are the primary reasons of the fluorescence blue shift; however, their lipid microenvironment must be also important in the initialization of the shift.
Solymosi
Katalin
K
Department of Plant Anatomy, Eötvös University, Pázmány P. sétány 1/C, Budapest, Hungary.
Smeller
László
L
Ryberg
Margareta
M
Sundqvist
Christer
C
Fidy
Judit
J
Böddi
Béla
B
D005453:Fluorescence; D007700:Kinetics; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000737:chemistry*; D010777:Photochemistry; D018515:Plant Leaves / Q000737:chemistry*; D011312:Pressure; D017434:Protein Structure, Tertiary; D011521:Protochlorophyllide / Q000737:chemistry*; D013057:Spectrum Analysis; D013696:Temperature; D020524:Thylakoids / Q000737:chemistry*; D014908:Triticum / Q000737:chemistry*
D011521:Protochlorophyllide; D044925:Oxidoreductases Acting on CH-CH Group Donors; C023886:protochlorophyllide reductase